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中議

口Ю

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Ｌ　貞質Ｖ

面螂　請

:ndLIStFia:

o!ogytechnology

Concepts. lu4icroorganisms are used in inCustrial micrcbiology and

biotechnologi' to create a wide variety of pr-oducts anC toassist in maintainrng and improving the environment

. Most work In industrial microbiology has been cai'ried out

using microorganisms isolatdd fi'om nature or modifiedthrough muiations. ln biotechnology, microcrganisms withspecific genetic characteristics Lan be constructed to meet

cesrred oblectives.

" i\4ost microorganisms have not been gro'"^/n or- described A

rnator chalienge in biotechnoiogy is to be abre to grow and

cl-rara.terlze -ihese observed but uncultured microorgan-isms rn ',",,hat is called "bioprospectit'r9."

. Forcecl e';oLuiion and adaptive mutatlons now are parl ofbrotechno ogy. These .:L De carried out in processes termeC" natural genai-i- engineerlng."

. The developmen' of meciia and specif ic conditions for theqro'.vth of microorganisms is a large part of industrial mi-

crobiology and biotechnology Mi-

一一■

crocrganisnrs can be grown in

controlJed environments rrith spe-

cific ltrnrtations to nraxinrize thesynihesis of desired Products.

. Microbial gro'',r'th in soils, waters,and other environments, whereconrplex nricrobial cornmunities al

ready are present, cannot be com-pletely contrclled, and it is not possible

to preosely define ltmititrg factot's orphysical condiiions.

. Microbial gr-or,'.th in ccntrolledenvironments is exPensir''e; it is

used to synthesrze high value microbial metabo-lites and other products for use in animal and

Biodegradation often can be facilitated by

changing environmental conditions.

Polychlorinated biphenyls (PCBs) are

u'iiiespread in<justrial conianlifl ants that

accunrulate in anaerobic river nluds- Although

reductir e dechlorinatirrn occurs under these

:.rnaL-r()hic cr.rndili(rns. ()\\ !€n is rcquired to

.()mpleie the dcr:radatitlrt process. In this

erperinrenl. nruds are being acrated to allou'

the final biodegradaticrn steps 1cl occur'

一ネ一

一．一、

一一一一一一ヽ〓Ｆ・〓

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||ギ 11

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ｌで■ダ

あ と 脅 〃

96,4 Chapter 42 lndustrial Microbiology and Biotechnology

human health. ln comparison, microbial growth in naturalenvironments usually does not involve the creation of spe-cific microbial products; microorganisms are used tc carryout lower-value environmental and agriculture-relatedprocesses.ln controlled growth systems, different products are synthesized during growth and after growth is completed.Most antibiotics are produced after the completion of ac

-: .' ,. ;.tive growth.Antibioiics and other micrcbral products continue toconti-ibute to anrmal and human welfare. l.lewer prod-ucis include anticancer Crugs. Comb!ratorial bioiogy is

making ii pcssible to produce hybrio antibiotics rnrithunique properties.The prociucts of industrrai microbrology rmpact ali aspectsof our Iives. These often are bulk chemicals that are used as

fooC supplements and acidifying agents. Other productsare used as biosurfactants and emulsificr-s in a r,,ide varietycf applicat;orrsDegradation is critical for understanding rnicrobial contri-butions to natural environments. The cherriicalstructure ofsubstrates and microbial conrmunity char.-.cter strcs piay rrn-portant roles in determining the fate ol chemicals. An;ero-bic degradation processes are rnrportant for the initialmocji{icatron of nrany conrpourds, especiaily thcse r.vitlchlorine and other halogenated functions Degradatroncan produce sirnpler or modified 6,,111,r6snds th;i rnay notbe less toxii than the original compoundSirseniors are unCergcing rapid de;,eloprrent, which'rlimited only by the advances that are occurring in rrolecular biology and other areas of sr ience. lt is nor,n, possible, es-pecially with streptavidin biotin-linked systerrs, to haveessentially real-tinre detectron o{ important pathogens.Gene arrays, based on recombinant Dl'JA technoiogy, allo,,vgene expJession to be rnonitored. These systenrs are berngused in the analysis of corrplex nricrobraisystems.Barteria, fungi, and viruses are increasingly employed as

bicpesticides, thus reducing dependence on chemicalpesiicides.Application of microorganisms and their technoiogy hasbotir positive and negative aspects. Possible broader impacts of applications of industrial rnicrobioloqy and brotech-nologl, should be considered in the application of thrsrapidiy expanding area.

'l'ltt nti<'robe will lnve

ndr-rstrial microbiologr and biotechrolr)cv both in-

" olt e the Llse tll- illcr,rr)r'Slni\nr\ ii) ir.hieve spccific

gotll、 、hctincr crctiting nc、、 prぃてluご 1、 、、ith lll(ヽ llclttrヽ aヽltlc or

illlprぃ 、:1lt thc cil、 irl‐ nillci1l IIntitト モ:itll l)liご lぃ 卜:ぃ 1● ■、 ti、 11 1,ti、

:itiditi● I]till卜 dc、 clて 、pcd focと ,、 ビ、、)I]I]iい dt'こ 1、 、tiChよ ヽP11ま ::ミ ltiCC t]―

[iぜ tll こnl1 11lcdical connPoし lnd、 (ar,Iibit)liご 、 11て「ヽnl ollcヽ 1l an、 一

fo「 :rlcd stcroids, solヽ iCnts organic acids.chcnlical lbcdstock、

422

anrino:rcids. anci enzrnles tilrit harc direct economic value. Thenricroorganisnrs cnrplorcd bv indLrstry hrre been isolated frorrnalLlre. end in rrurnr clr\c\. \\crc rrrodified Lrsins classic nrulation-sclcttion pr oceclurcs.

Ilioteehnolog', hu: l Iong historl. rnvr-rlving ti.ie irsc of liling or-ulrrrisnr: 1'oi' conrnrer-ciiil hcncf-i1. In thc lasl \,'\ eral decacles. the

tL'rnr hlr\ i-lacr-rnre usstlciltted u ith tire trOcliiicltiitrn ()l' ltliLror)r

sairisms thloirgir tht- usc o1'ntoleculur hiologr. including the use

irl recornirinlLnt D\,\ Iechn()l()g\ l.:ta <lrultttr l+). It is no\\'pos-:ihic to nlinipLrllrtc r.nL-tic inf'ornlrlion rrntl rlesign Ilroducts such

as pro{e ins. or Lo nrorlilr rnicr-obial gcnc e\prcs\ion. In iicltlilion.gcnciic inl'or-rnlilion e lin be ir-rrnsicIred b.'tu e eir nrerkedlr difler-e nl !r-oup\ ril'or'!lnirnrs. r,uch lrr hcluecn llictt'r-ia anil plants.

l-irc us.: ot' nricroorglinisnrs in industriai nrir:irlbii'rlir,cr, and

biotr'e hntrlogi lollrrn s li logiclil \cLiLre nca. Ii is irecessarv first torrltntrir ()r' ! r'tlrlrt lt nticroorgllti:nr thlil clilric: out lire clesir-cti

[]r'1)LL'\\ in tire rrrosl tll'ieient rlllinrcr.l-his nric|o,rrgenisi| iiren isii\cil. cither in.i coirtlollcd r-nvir{)nnlcrrt:rich lrs rr i'ernrentrrr-oritt c,rntplcx niitullll \,\ \tL'nls. sttclt lis in srtils or \\ ll[r'rs. 10 lchiu\ e

.irtiilic Srri j..

CH00SiNG A/1iCROORGANiSMSFORINDUSTR:AL MiCROBIOLOGYAND B10TECHNOLOGY

「 llc iir、 (ta、 kl()l(tn i11(l tl、 tritti l1licl obiol()ti、 1 1、 1()llin(1:1、 Lli t(lblc

11licr()o「 tani、 11l olnC that is tc1lclictlH、 ‐ 、lti卜 lc ctt、 、 to l〕 1[lilltt、 ill

(lndゞ |● ｀` tinCiヽ、cll、 tlitc(11()l c、 (l tt(■ it)nt,1 、cIっ alを 1li、 )1〕 ()十 (lc、 1lc(1

1)|()dtiCl、 ,へ 、、iclc 、r tl「ict卜 ol tll(Crl〕 ati、 c al)l)lt)を iC1lC、 (Hct,ヽ ailabic

I[11〕 ■int fr()n、 i、 ()l■ tiini n3ic,,()(01■ tlll iヽ 111、 1'1(ヽ 11l ihC C11、 irolllllCnt t(う

u、 inぎ 、●1っIni、 tictttc(1 121()lccuLtr tccil:li(luc、 t()l11()(11「). tlll c、 iヽ ling

nnlcI()()1 2tti,lヽ 1〕 ]

Finding Lttici′ ooroattiSms tn ttatttre

tinlll rclati、 cI卜 「cccnti卜 thc 11ltl(1、 ()urcc、 、)F nlicI()bittl culturcs

lol‐ t卜 c i1l i1ldtl、 trial 11licI(♪ biol()t、 、、CrC llttttlltll 11〕 11lcl iclis、 uch tt、

、oil 、ttinlPlc、 、、latcr、 そ11ld、 P(,ilcd卜「c tl(l a11(111‐ tli((｀ tllturc、 1l ol“

aH tllca、 of thc、、orld contillしlc lo bc cヽ そinlincd lo idclltilヽ 1ヽ 11[1111ヽ

、、itll clc、 il t、 1ぅ lc charactcli、 tics llntcrc、 t in huntint f()r nc、 1111-

croolga,11、 1111、 、ol biol)rospcctillg,colltilluc、 loda)

Bccau、 c olll)a lllinor Portion()f the nlicrobial spccics csti―

n]atcd to cxist il〕 11loSt CllViI()nnlcnts has bccll is()latcd or culturcd

(table 42.1)tilclc is tl continuillg cffol■ thl()ughotlt thc、vorld to

gro、、 lincsc しlnctllttlrcd・ 11l icl()o「 gtlnis11ls 1ln tCrllls of thc llll101-

c11、 ir()nnlcnt、 11l o、 i nliClobc、 that can bc obscr、 cd ha、 .cn()t bccil

culitl「 c(1(,「 icicntilヽ c(1(table 42.2, ヽヽ:ith i11ビ

l ctlヽ CCl ill(crc、 t in llll

cIぃ いitil di、 cl、 1[〉 Iュ liご「()卜 itll cc()log) an(l c、 Pccia‖ ) i11 1lnicrooI

gani、 1ぃ 、 1l olll cヽ tl C 11lc c ilヽ i10n:1lcnt、 {Tcchniques &rゝ pplications 42.1). 1llicI()biologist、 arc lcarning hO、 lo gro、ハ「

964 Chapter 42 lndustrial Microbiology and Biotechnology

human health in comparisOn′ microbial groぃノth in naturalenvironments usua‖ y does not involve the creation Of spe―

cific microbial productsi nnicroorganisms are used tO carry

out lo、ヘノer Value environmental and a9riculture― relatedprOcesses

O in contro‖ ed groν、/th systems, diferent products are syn―

thesized during gro、へ〆th and after 9ro、へ/th iS cOnnpleted

4ヽost antibiotics are produced after the completion of ac―

tive gro、へ′th i ′ 二 ′・ Antib10tics and other naicrobial produ(ts continue to

ilよ留l:∴ 11『 :infrl」∬誂∬|:鷺 rti:ふ島ニmaking it possible to produce hybrid antibiotics ν、/ithunique properties

e The products of industrial micrρ biology impact a l aspects

of ourlives These Often are bu kと hem calsthat are used asfood supplements and acidifゞ ing agents other prOductsare used as biosurfactants and elη uls fiers in aぃノide varietyof applcat ons ヽ ′

・ De9radatiOn is crit cal for understandi∩9 nni(robial cOntri―

butions tO natural env ronrnents The chelnicalstructure of

substrates and micrOblal cOnnm_ln ty cl‐ 8raCter st cs play,fn―

pOrta∩ t rolesin detertnining the fate_lfchenn cals An3ero―

bi( de9radation prOcesses are inlportarnt for the in tial

modification of nnany colη POul:dS′ especia y thOse 、4ュ thchlo「 ∩e a∩ d other halogenated ful¬ ct Orns De9radatior¬

car]produce simpler or n¬ Od fied cゥ nI「 OundS that lTay lnOtbe less t,xic thall the ori9inal COmpOund

・ BIosc n,Ors are under9oing rapid develoll1lnent′ 、Aノ hich Fli「nited only by the advances tinat are occurr r¬ g in iη Olecu―

lar biology and other areas of science ltisinOぃy possible′ es―

pe(ia‖ y ぃノith Streptavidin― biotin inked systems′ to haveessernt a‖ y rea tinne detection Ofi:mportant pathOgens

e Gene arrays′ based on recombinant DNA techno109y′ a lo、ハy

99'C eXtrF:ユ 1911 19 be mOn,tOled These systcms are be ngused in the al‐ alys s Of cOnlplex n)icrobial systenns

・ Bacteria′ fun9i′ and viruses are irncreasin91y er〕 np10yed asbiopesticides′ thus reducing depe∩ dence Oin chenn calpesticides

・ Applicat On of mlcrOOrgan sms and tino r techno10gy hasboth pOs tive and negative aspects Possible brOader im―pacts Of app ications ofindustrial:microbiolo9y and biOtech―

1¬ olo9y shOuld be cOnsidered il‐ the app ication Of thisrapidly expanding area

422

arrrino aciils. an(l enr,\ ri)es thrit hlir r- direct econitnric l.alue. Thenricroorganisnts ernplorcd bv industn hare bcen isolated frontniltr.lie. and in nlin_r cil\as. \\ crc nrociilied using classic rnutation-srlt-'et ron procecl ures.

lJiot,-chnt;lo"r hus li long histun. lnr,c.;lr ing iite irse of lilin-e or-!iittisttts foi con.rntr'rciril hcnef-i1. In the last stvcral dccacles. thete

'nr hlrs bccr-.n)e uss.ciete d u ith tire m.rirf ication of nricroor-

qanisnts thr.ouglt the rr:e r-rl'niolecular hiolrtgy. includins the useo1 r-cconibinunt D\.\ lechnolLrgl (tt,e t ltttltrrr /4). It is no\\r pos-sjblr io nlrrriprrlute g.nelic intorntrlion rinri desi!,n products sucha\ pr(itcin\. or to tnodilr ntretlbilil gcne e\pression. in aclciitiitn.gcll.lie ittliit tttrttirtp tltp bc translc;rerj br'11ren ilarkedly clilfcr-ent grouit\ o1 orLlinisnrs. ,.uch tis hetu ce n 1:lre lcril and plants.'I-lri:,sr .i,ticr..r!u'tis,ts i, i.dLrstr.iui l,icr-,bi,Jloc_r, anclitiotte hrtrrltrli lolirin s a louicitl scqLlcnci.r. lt is ne .e:srr1 Irrst iordr'ntil'r {)r.!lairlo lr nricloor!lrnis111 111,,1 cer-r-ic: out rhe desircdllr'(rLc\\ in lhc nrosl clf icie nt illllnndr. l-his nricroorg.rnisnr iiren isu\rrrl- cilhc-r in ri r'ont|0]lccl rnVrronnrent sucir as 3 ierntenlcr 0rtttirr111|li'1 niltLiJel s,\\tclns.sLrihlsinsrtil:or\\alcrs.tlrltchicr,e.1te.if ir'Sir:rI.

CH00SiNG A/11⊂R00RGANiSA/1SFORINDUSTR:AL MiCROBIOLOGYAND BIOTECHNOLOGY

‐「 |〕 c lir、 1[を 1、 k lol(ln il〕 (i tl、 tritll illicrol)101()=i、 ( 1、 lo l｀ illdをて、tlittll)lc

ill ic「 (,o「galli、 l11 01RC thtit i、 gcllctica‖ )、 ltiblC Ca、 卜 [(D l]lai11ltlin

tinci=r(,、 `al〕 d、、cH、 tlitctl l()1 じ、〔laclion()1‐ 、cPttl 11(loll ol(lc、 1lcd

l):('CI tlCl、 :ヽ 、、1こIc 、aricl) (、 1(11(Crntlll、 c tll)lD:()を lChCヽ alc a、 1liltllDIc

ra11ビ i:1= f10111 1、 01:lti l〕 ゞ1lliCI()()1‐gani、 111、 11()111 lhc cnヽ il()[lnlcnt t()

し1、 in= 、01)lliヽ tictltcd 111()lcctlitll tcchllicluc、 10 nl()cI‖ 'ヽ all Cヽ i、 ling

nllcI()。【tanlヽ 111

Finding MicrOOrsaniSItts in ttatttre

tlntil rciati、 cI)rcccntl)lhc ml(1ヽ 凛lcビ 0`1 micRbiJ culmrc、lk:)I tl、 c in in(ltl、 モlial l〕 nicr01っ i(,loご )、 で`「C Intlltll tll lllalcl ial、 stlch aヽ

、()il、とllnIDlCヽ ヽヽla(cr、 and、 Poilcd lDlCを 1(1を 11ld ll■ li((「 tllturc、 lTonl

[Jl alcti、 (「)li thc、、01ld c01nti11し ic l()hc c、 a1llillc(itO i(lclltifv、 (1を li11、

、it11(lcsiI(1卜 lc ch(1lactcli、 tic、 Intcrc、 l in litintillЧ ll,I‐ nc、、‐1ll i

croolg:lnisi11卜 、ol bioprospectillg,cOntil〕 tl c、 10(la)

Bccatl、 cO:11)a11linor portion()f the llnicrobial specics esti―

1lnatcd to cxist in lllo、 l cnvil‐ ollnlcnts has bccn isolatcd()1‐ culturcd

(table 42.1).tllcrc is a contintlillg cllbl■ throughout tlle、 ハ「orld to

gro、、thcsc u1lcul(tll cd｀ 11lic100「 ganislll、 11l tcr11ls()f thc lll[l10r

C llヽ 110nIユ lcnt、 n]()、 l nlicrObc、 that call bc(♪ bscr、 cd lla、 .c ilol bccn

culttll cti()l iticlniliヽ c(1(tal)le 42.2, ヽヽ=ith i11ご「c tlヽ Cd intcrc、 t in ilni―

cII)1う itil di、 cr、 iぃ 11liC10卜 itll cc(、 lo=) tt11(l C、 pccia‖ 卜 i11 1〕 ]icroor―

gュ 1〕 1、 in、 「1()1〕1 ビヽ tl cnlC cn、 11‐oll11lCntS (ltchniques &

へpplications 42.1, 1〕 nicit)biOIogist、 arc lcarning ho、 lo gro、 '

′ノ

／

7功ピ777'こ ′て177ピ ″J〃 /2aν

`″ι`Za,1

ndustrial nlicrobiol()g)and hi(、 tcch:1()10ピ ゝ‐ b01h in―

L/ '.'olr e the usc of rnicroorglinisnts itr lchier e .pcciiic-!oalS. it he ther= crcliiin-q na\\ itr.oalLtLt\ \,, tth nt()netai'r, r 11111.. ,11.

improrinS ihe environment. Indu:rrial nticrobiolocr. as it hasIraditir)nallv der eloped. t-ocuses on prL)ducr\ \uch as phai-ntaceu_tical and medical compounds iantibiotics. hormones. trans-formed steroids). sol\rents. organic acids. chemical feedstocks.

423 Choosing Microorgan isms for lndustria I Microbiology and Biotechnology

ilic Archaea in Biotechnol

There is -rreat inierest jn the characteristics of archaea isoiatc-tl fronrthe outllo'rv nriring regions abovc deep hvdrothermai vents ihai re

iease rirter at 250 to 3-50"C. This rs because these hartlv organisnrscan -gro\\r at ierrpeiatuie-s as high as l2l "C. The probhnts rn sr{,u ingthese microorganisnrs in a laboratory are formii]able. Fcir examnie. tirgrou,some o1'them, it is necessan,to use spcciai culturing chanrbcrs

and other specralized equipment to rneintarn ulier rn the liquiil state

at these high temperatures.

Such rnicrrrrrrganisnrs. terrned hr,pefi hernrophiles. u ith optilnurnlrou'th temperatures of 80-C or abuve (.rr:c pp. )2-i-l).1t. eonlrLrnitunique chrlierrges in nrtrient acqursition. nrctrrbolisni. nucleic acjtirepircation. and grorvth. \.1anr oi.these a;-e inaerobls that dc-pend on

cle ntental sullir tr;- 1in ic ion as oridants. Enzt'nte stabilit;,, is critical.Scinre DN.,\ llol\i.n.rases are inherentlv stable at 140'C- q,hereas

nranl' other erlz\'rllcs i.iie stabilized in r,rYo wjth unieue therrropro-ttct3nts. \4hen thcse iinzlirnes are separated fiont their proteatant,ihev krse their unique iherrrrtrstebilitl,.

Thesr cnzrnrrs rrra., have intportani applications in mernlne prl-cluctjon. nreiai leaching and rel-over,,. and in irnrnobilizeC enzyme sys-

lrnrs. in atidrlroir. the prissibi litr til'selectivc stereochentical mtxlifi cationrtl conr-Lrltntls noiliallr not in solutiort al Ltrt'cr tempcratures inay prr)

\ i(lL- n!'\\ r(ruic: firr directed chernical sYnthcses. This is lut excitrng alde rirrLnrlinl arell !11 th. nrrxlcnt biololicai science: to nhich environnttit-tel nricrobrol0gjsl: cul.r riluke .ignitlcaltt contnLrutions.

Estimated Total and Known Speciesfrom Different Microbial Grou

E sti m atesin V;:ious

of the Percent "Cultured" MicroorganismE nviron ments

EstirrratedI'otal Specirs

Knou n

S pecies''I)erce n !

Knorr n

[-nr ironnte^1 f-sl irnatcd I)crcent Cuitured(iroup

Viru ses

.Archaea

B rLc teri:r

Fun-ei

130000b

40 01Xlト

150003060000

5000<500480069000400()0

14]ビ

1121

う

67

Seawaier

Fresh q'ater

Mesotrophic lake

Llnpolluted esiuarine s,arers

Actir,ated sludgeSediments

Soil

()001-0100

02501-1001-30

_11_15 _′

02503‐

'Mid-1990 values and sh()uld be rncrcf,s€d li) 50!7.hThese values re sub\untiail\ uildereslimnred. p€rhaps b) l-l ordr.\ oi rnaqniludc

'l I indicales fiat these yalues ee probahl\'!!ro\s overeslimatei.dSmall subunl (SSU) rI{NA dak indicxre rnu.h hrsher in \nu di!enrr} dd Si.cn h\ cukure h:rrd sludi.\

Adapledfrom:D.A Cos3n 1000 \'ri.robi!l genonr\-theuntafrprdresourr fiSrr,cli Il:lJ-l6Table l, p. 15.

illcsc Prc、 1()tl、 l卜 ttllctJitllcti 11licIぃ bc、 11]、 ,iticiil〕 ゞthc Llヽ Ct)1)ヽ i11-

glc― ccH =cl 11liCrぃ c11ご 〔lI)｀ tll(ltiぃ Il を11)IP「 いtiChC｀ lhtil llllぃ 、、 :ltill iて ,111

(11■'tlsi()1l alltl nRicrt,bitll さぃ:]lll]t‖ liCatiぃ n 、、1lilc :1〕 illinli/iln g 、)、 'cl―

gro、、ith b) 11〕 o「C rtllDill―

=「

o、、ing c、 )11]I)Ctitい I、 1 ‐ ‐「 1 11‐

Genetic Manipulation of rh/1icrOOrganisms

Gcnctic lllaniplll(lti(Dn、 arc u、cd lo producc i〕 ]icF()Orgallis11ls、 1ヽ111

nc、v and dcsirablc char[lctcristics Thc clas、 ical nlcthods of llll―

cI()bial gcnctic、 (、 どで'(/?て り,′て,′ ノマI Pl(1)tl、 ittil rt)ic in tI〕 c dc、 ビIぃ P―

11lCl11 01'ctlittlrc、 lbr irldヒ lstritiI 11〕 icrobi、 ):ぃ=、

MutationO1lcc a prollli、 illビ ごultulc i、「 しlnd [1、 Liriピ l卜 ぃi tcCi〕 n itltiき ごヽti:う 卜C

uscd for culturc illlpro、 cnncnt illciLItiillご ご1lCrllictil l■ utagcn、 [lnd

Source: D A Coq in l{I\). \4 i.r ot,ial _cenonres-the ur lapped resource. Ibrec, I 8: I 4- I 6. Tsble 2,p. l5

tlltiti、 1()lci lighiぃてて■ノた1/Pr`′ ′ ノノI A、 a llじ 、arllplc.(1lc ril、 t culttllて

ぃ1 /'で ,″ ′(′ ///″ ″77■ ,「 tl子 |″ ,1 ヽヽh i t111(■ tll(l bc tlo、、11 01Rl、 tllldCrヽ tati

C01ltliti(、 11、 )iCl(lcci l●、、 tt,I]CC1ltl tlti()11、 ol PCnicillil]In 1913

、lrain o「 /t,′ ,ltア /ル ′`′

″ (カ ハ、て,マて'′ 7'′′7'ヽ aヽS iSOlatcd― straill NRRI

1951 -― 、、1lich 、、tl、 lullhcl in]PI o、 cd thlough lnutation(「Igur

12.1, ‐「 oda)ilno、 t pcllici1lill l、 lD1 0(luccd、 、.ith et′ ガ(1′ ′′′

“′7,(力 ′]ヽι

マて'′ i!`′ ′? gi■ )ヽ、n il](lcIぃ tDiCヽ tilrcd iモ 111lcntcr、 、、hich gi、 ics 55-1'ol

highcI PclliCillil〕 卜icld、 thtln thc origillal static culturcs

Protoplast Fusion

I)rotoplast ftlsioll i、 Il()ヽ ヽ ヽヽidcl〉 t.、 cd、 ith)casts and lnold`

1ヽ● 、1 01 lhc、 cl]liご iい い1=tl I]i｀ 111、 tll C.lヽ Cヽ tltll()r()f a sirlglc lllatinl

モ)1・せ ｀`hiCI(1ご こ:き ょ、きヽ 1:]ビ ごl‐ttilぜ こいi l ttinti()111 11ltlttltioll、 lh〔it coul(

lct`」 1(1、 ti t:ill tit=さ ilcl ti:い い Iて こヽa rl、 く,tit tcnctic、 tudic、 ヽヽith thcsl

1111ご [、 1● Iピ ti「 i、 111、 1■

1 1ヽ1 li〕 i tt、 1、 tii C PrCPtirCd b)i gro、 ving tllc ccll、 il

an ist)lonic、 oltlllon、 hilc trcatinゞ thcl11、 ‐ヽith cnzylllcs includinl

16 Chapter 42 lndustriai lr4icrobiology and Biotechnology

NRRL 1951[1201

1

NRRL1951 B 25[250]

|ド

X-1612[500]

|と・wis Q176〔 900]

|:

Bし3-D10

Pt:nit'iliiunt rrtqtte.fttrrii have been l'used r"'ith those of P' cltrt'so-

qettiun. Even veast proitlplast-r and ervthroc\'{es can Lre fused'

Insertion of Short DNA Sequences

Sho|t itngths ol chcnricallY si'tlihe siz':tl I)\'\ seqLrcnces catl be

inse rlcd intii re cipicnt qlicro(lrgenl.5111s hr tlle lltirce ss o1' stte-

directecl nrutagenesis. l'his can crciitc \lllllll gcilcllc ltltel lltign.''

lcaciinglitachiurgr\)1'Olle()iscrellt] atllitloltcirlsil)lhcIal('L'tplo

te in. Sirch ntin0r antilto liciC chrngcs itare br'rll lilLrtl'-1 itl lclld- in

lnan\ cuscs. to uile \Pcctcii ciletlgcs iil prolt'l;l chlrr:ritcristics' antj

itar.e t.esulietl in nc$ prodLlcts -sucit :i:; Irlilie cilr it'trnttlctllltllr re

sisl:iltt erlz) tllels atlti e llz\ lllr-\ thlit clin clitltir zc tle srred reactiot]s'-l-he'c apprortches ltrc ilal.t o1 tilt f ie 1tl o1' protein engineering'

49133P,23q

48-1655

49-482

49-2695

60-529

50-1247[1.506〕

48-786

|

i tl

48-1372[1,3431

1 ‐i‐

49-901

49-24291 Fi

50-724

50-1583

1 :'1,

51-825

1‐52-85

11■ |

52-817

11■53-174

:|‐ ||

53-844〔1,846]

llc,r.,are industrial microbiology and biotechnology sinrilar and

different?What approach is being used to increase the recovery of pre-

viously "uncultured" micrcorganisms from the environ'

i'n e nt7

What rs protopiast fusion and what iypes cf microorganisms

orr UsPi ir Inis orcr.::lDesci'ibe site-directeci mutagenesls and how i1 is us:d in

biotechnoiogy.

!!lat is protein engineerino?

3

)-2166

50-25

0-935

茸＼、

53-399 53-414

12,658] [2.580〕

馴fm ttf∞

馴翼

F

51-20A2 51-2033 F3121140]

F3‐ 64

12.4931

'igure 42.1 \Iutation \I:rkes It Possible to Increase:'ernrentali<)IrYieltls. A"gcncalogr'oi'thcttttltlLtirrlll]1 |LL'\\L'\

rsetl i0 inurcasc pc-nicilltn Iieltls $ith PdtitilliLtttl (l1t\'\()!(tltttlt

tsi.g X-t:uy,treilt.te.t (X). U\r trcat,iL-nt (Llyt. allcl llltlstard Sll\

N t. BY use 01,thesc lllulatioltal processes. the ficlci lr as itlcreas.'.1

l.ont 120 lnternarional L.lnits (lUtto 2.-580 iLl. a l0-1'old itlcrease .

-.innrarketl transfers u'ere usecl for nlutallt cro\\'th alld isolation.

fields in international units/mi in brackets'

_.ellullsc and heta galiictr,rrr,nidase. The p!otoplasts are therl rt'-

rtncr-atetl using osrnotic stabilizers sttch as sLlcrose Alter rcge n-

.:r-ririrrtt rtf the ccil ri all. rhe neu Prr:ttlplaslll 1'ttsitln prodttcl clltl tte

ir'eJ in furthcr studies.

A rIa.lrtr adr lntage ()1' the protoplast f'ustorl tecltlltclLrc i. thlt

protoplasts ot'dillcrent rlricrobial species cu be fused' er en il thcr

,ra nu, closelr lirlked tarontlllrcall.v-. For cxatrlple' protr-lpla:ts oi'

Transfer of Genetic lnformationbetween Different OrganismsNcr,, rrltcilltitrr, i.iare lrir,:n throLrgll tilc liilllslL'l ,rl nLlcle ii.lci.ls

l-..'1..'.rrtrtii1l'ercrtt (it !l1llt\ll1\. uliitll ir Irill'[ ()i tirt'rlipirllr 11t'r't'lrll'l

ing l ie1t1 Lrl ettntbinttorial hioloul t tablt -ll'-1 r l his inr trlr et tire

,r,r,.,.1a, r)t' !f It!-\ fl'ttttl tlll.' ot-latlisttt it-tto rllolhcl" gir rllg tile lt

i:iprr':tt rltitctl iellltitilitl.': .tr.li lt' ltll itltlce'cti ' l1t'ltitr t" 'lllrr,riti lrt.ltri.ltlilr,il de!tlt,l.ttl\rjl .\l llll|r)11"1 l '''ll\ c\iilllplc (11'tlli\

ap].,',rach \ a: thc rl ailil{llr (r1' th- 'stipcrbu!" paicnte tl b\

{ \1. f llikrrlribat-tr rtl l(lli. ri irich hrid itrl lllri'll\c(l caputrilih o1'

1,r,11,1.111irct1 .it!tltti;-itirltl. Th.'-lctle: lirt liiltihlotic plrltlLtcltoll call

lre lLl:l:1'L:l'letl irr:l itlliirrt'l-!lLilitllt l]llti lllLrtiriie::tilrrlllcrllliihi()ti!''r)1 C\tn ti) ii Iloil;11111f!ir111'-lllrl,lrieitl-r illiclt'L't-!'tltt'trl othct er

Itttlllie s ilrc tllc e\fi'\\i()n' tl !- ' "lt ()l ihL'

'll1) lllr !lt'11illrill't\'

tl-rrnr l)ir'rrrlr,rttt)tt(t.\ Pttiitltt lrird tltc ptiltlLtetirrtt o1 Pccliocill' a blc

tc'r-i0e tlt. ill a r cast ust'tl itl lr tl'tt' l'ertltelltltti0tl t" 't ihe Prtt lilre It

aor'tlr()il i n !' i.ltcteriel aolrl,tl I I I Illlllt \'D\.\ erpr.es:ioll tn tlitlcte-nI I rl--]llrll\lll\ Lilll llllPl()\t lltrlduelitltl

elllcicncr and ntinintize thc Pr-rnllu.rtir rtl rte P\ rcquiretl bctorc' the

prtltiuct is reat1l lbr r-rse , For cralllple. rectlttthtnant hltcultlr it.uses'(.\c(

lp. 10'1 5) can be repliclited in insect ia:-rae to achiere rapid

l:u s.' sctlc prrtiuclion of .L de :irctl r irui rrr pl ote i ;t. Trltnsuenic plants

r.1i,.71.r, i.,.,';'..'l', -i-':il1.r\hr'llt("11r)ll'.ilil'lr!liriilrl:j(qu'rtrl;-i 1.. ,i : ..:i .i\ ,.1 i;..:.:r..:. :'t ,.ii;.i. \lli"'' ll:l.L:;1.ll r' ''r :r' I !rl-

Llrll!ll irt '-. ';5i. f)\.\ :,li,.r l: Iri-lili i' irr 'lillllr 'll'rrrt 1l lil r'i\lil-l

D\ \ err:rtL'ri illiirt'ilt,'1:.l,la' ;iil!i il :lini'-!r'lil r\('i 'i ' 11'1i I t tl:'

.\,,rrtlr.;:tn!3 i)I ga!lal1! iill'trl'illelt,ln il15r) ai1ll ha ill5'l-i'Ll lntO

nlie fr)organisttls li:ill! \ iat(i1'\ lnd re conlbitlalll DN.{ tecllniqtir's.

47-638[9801 47-762

|

47-1564[1,357]

5120P,521]

|ド>＼、、、

52-318

' 嗜可~~¬

52-1087 1

425 Choosing MicroOrganisms fOr lndllstrialヽ 4icrobiolo9y and Bictechnolo9y

Property or Prodtict′I■ansferred

～licroorganism tised cOmbinatoriaI Pr∝ ess

Ethanol production Esc!rc rit ltia coli

Combinatorial Biologyin Other Organisrns to

1.3 Prooanedioi prociucrion L.. coti

Cephaltrsponn precursor Penicilliun tltnsoqenutnsv nthesi s

Lactic acid prrrCuitionXf iitoJ production

flrcatini n aseb

Perii oci n'

.{celone and butanolproti uct iitn

in Biotechnology: The Expression of Geneslmprove Processes and products

'\tttchurottt.tce.s t ereti-slae A muscle bovile lactate dehydrogenase gene (l-DH A).expressetJ in S. ceret,isiae..\. ceret'i:iae 957r xvlitol ccnversion from xylose u,iis obiained bt. irarsfr,rntins th; XyLI cene of plclla

rlrpilr enr'odin: a \r lose ,.J,u,,,." i",, a. , , "r,,;,,, "

,,,f ,,,, iUi. ,, .rrir,,, ,rl,],.i.,,,orgattistlt lbr the protiuction o1'rylito1. u,hich servcs as a sweetener in the tbod in6ustrr,.

E' crtli Expression of the creatininase gene fionr P.seutLtmotut.,tT;aiilri R:65. Cese inserted *,rth apt-rCl8 vechr. .,'

lntegratron tlf pyruvatc decarboxi'lase iind alcohol dchvtirosenl:c ll from Zyntomonus ttutbi! i.s.Introducrion of g:nes rrom the Klehsitllu pncutttortile ait re-gion into E. r.ctli rnltde pcssible

:rnaerobic 1.3-propanediol productiolt_Production 7-ADC rld 7-ADCA' precursors b1 incorp,ration rrf the expandase gene of

Ce phalosoyxt rin ac rentatiutrt itto penir i I I iutn by transfcx-ntarrun.

lirpression of bzrcteriocin t-ront PeLliotot.r;u.v ut itjilutriii in.!. rez,r,i.iiae to j;rhibit u,ineconlantinan ts.

Intrutjuctir'rt rrl; .hrrrrle \c\'r,,r r''1,, L.,tL.t I,t,tt',rr, /,,,. r, r,r rJrfr,,\(J (lrrrrl,lrilrrslufnr.,ironprotocoi. rvhrch resultecl i n ;crr,,nr- rnij bui.-r nLrl lL,r r l.ju!,n.

S. i ereyi.siue

C Iost ridium d( (tobutyI i( utn

一

，

守

「

ACA=?aminmephrlosfomnicacid:7,ADCA=Taminodecacetoxyccphalosporonicacid

t.t,.iue. lu:t ljio-l7 {5h \r1v'f i,JUr

Adaplcd from S ostcrsaa劇 :L01s、 Onland J Nieison 2(〕〕,MctabO:ic cnginccin8 of Sα εε′r7r77・ ●`′

∫ε“でドなた′ピ MicrObiol MOI Bi01 Rev 64(1た 34-50

\ e ctoi's l.ttt, scr'tirttr i J.-it jnclrrilr artif icial chr()ll()-\()ntL.s \ri.h asilrosc 1'or' \eltst\ (\,rCs). [rac.icr.ilL (BACsl. I)l bacleriophage,tit'i r c'd t lli'r)al.soire s (l),\cs ). a'd ,*1rlr,ilia, a.lif icial e hr,,nr()\( inlr\ (I,1.A(-r ). )'AC's ilre csl)ecilill\' \ arlLrahle because liir!cD\-.\ scqLren.c:; 1()\er l0(l ktr) (.a, Lre ,laint.inr.rl in the \.\f.ris I\eillltsle ait11rlnor0ntr tjt \ L,tiSt Ce ils. ,{ g0ttd erlr1lnle 0l 1gg1111.

Lr\. l\ Jlro\ idctl hr tire r lrus that causcs 1'oor,anr1 rlloulir r-liscascof iatilc anrl other liiestock Ge nctic jltlorrrlrticln tbr u lirot-and1l()Lrlli \1j\a t\c r irLrs untisCn calt hc incol.ir()trtled inlo l.,. r oli. l'ol-liirr ed t.r lirc r'rPreriron .i'this gcnctic infirrnmtic;n and sr.nthrsiso1'tlte .gent, Itr()(lLltl t iot i-rsc in vaccine Ilr()duction (figure .12.2 t.

(icnt-tie iitt.ritr;rlirrit tra,s1'cr all.*s thc. ltr.ducli,lt rr1. spe.i1r. llrotdin\ rinrl pr'ptide -s u ith.ut contJnrinarion bt siinilarirr.tlucr: thut nrrghr hc sr nrhesizttl in thc original or-sirnisrl). l-hisliPp,.rrrr1f1 aiilt dL.arr,.rsc lhc lintc lutd c(ist 01'rCc<trerinl and puriir rrg u Jrotlrrcl ..\nrtthcr ilta j()r t(l\ ailtitge ol'peJrlrde prodLlclionu rtir i-riotechnologr is tirrt oirl_i Lrioiogicallv acti\ c stcrc()is()rllcl-sr1r'c llr()duc.d. t his :Pcciticitv is rLrr-luired to avoid the possibleharrrlirl sirle eflecrs ()i'ir.cti\,e stcreoisonrers. as ()ccLlrrecl in thethal idontide clisasler.

What is combinatorial biology and what is the basic approachus€d in this technique? What types of major products havebeen created using combinatorial biology?Wh,v mioht one !.!/ant to insert a gene in a foreign cell and howis this done?\l/hy is tt i,nporiant to produce specific isomers of prcducts foruse in animal and human health?

Modification of Gene ExpressionIn ad,-litirirt trt inse rtrn! lc\\ ,jL-nc\ rn .rj:u1t.nts. it ltlso is ltLr:siblt to ntodilr gL,ltc iaqilllt jon br chalginr, ,,cne tr.in.ijl ll)tiirn. lu:ing llr()teins. r|r:litin!, hr br itl llr()nr()iers. lrnd re rnor in!1'ccrlbirck regullition r.()ntt ols. I hc\c liIlll()itr.llc\ ntekc it lto:.:ihic ltt or e rprodLtcc u rr ir.it r lir.iclr o1' 1trotlircr:. lis sho.,r n intable 42.-1.

l'he ,taclil'icatiiilt.1 gc,c r\Jlr-c\\i.n .rls. i;i. itc usetl [. i.icniiona]lralternrellltrolicpiith\\irrsb-t in.reirrationorder-eLulllronof'.[.1-!iljc !]unr-\. ',', hrulr is tlte f'ie ld o1' pathtar architecture. I-,n-clerstanding pathu'a! llrchitcLrlLrre rriuk:s il J)ossihlc 1O rlrsiln apalh\\'av that u'ill be ntitst ell'icic'nt hr rrrotciin_q sloriet-()r en.rg.ticallr niore costl\ routrs. -l'his arrpr.riiir hri: hecn urerl 1o ir.rpr.rcpenici J lin prorluction bv rretabolic pathu a-r €ngineering (\l pl._ r.

An illteresting cler elopntent in ntodil'r irtr Se rrr c\l)l-r\\lur.u,hich illustmtcs metabolic control rngincering. i, th;rt ot tlrcrittg c()ntrols lOt the svnthcsi\ ()1' l) copene . an jJIpr)riJitt iiltttr.,\ldant norrnnll)' present at hich ier cls in Iorratoe s anci tonrrrttrproducts. In this case. iin enqineerctl rr.gulalon circuit u a, clc

signed tit control lvcopene s\,nthe-\is in response to the inte rn.ilmetabolic state of E. coli. Another reccnt derclopntent is the u-se

ol modifled sene expression to produce varianrs oi rhe antibioricerythronrycin. Blocking specific biocherrical steps (figure 42.3)in pathu'avs for the sr nthesis ol- an :tntihiolic precursor reqr-rltedin rnoilil-ied final prrtrlucts. l-hcse rrite ; eti pr.rJLrct:. ri hi,:h hiir.-slichtl-r'ditlercnt strriclrlre\. ealt be ir,irJ irrr lheir p,r::ihlc :rt:-tinticrobial cftects. In ailditiirr. ir,i ihe Lr.c .i rhis appririi.h. ir i:p()ssible to better establish thc \rnrcture lltrctii.rn rrlatir-rnships ot'antibiotics.

VP#1protein

Foot-and-mouthdisease virus

Viral RNA

4

Viralproteins

Beversetranscription

Clone oi

ごVira DNA wlh

VP,1 9ene

謗¨I RettrctOn

l enZyme

r繹『鰤中

1

|

▼

不 :謂献

ll習∝mttOn d

Ａ

ａ‐ＲＮ円（ノ／ｔ、．ノ／

Ｖｉ

ｆ。

Foreign gene

Bacterialchror]]osorrte

ヽヽ＼＼ヽ、

VP11

protein

recomb nanttl acte ri a

Figure 42.2 Reeornllinant \accine Protit-tcti<ttl'

( r1' rer()tithi Itlltll D\.'\ 1c'cllrl tqLte:. I firrrl litltl t tlr itlih

aoaGa

30eVP#1 protein from recombinant bacteria{or use in vaccine Production

( itni. cotlinl tor ile:ircrl Jtroditcts can be e \Jrresserl in dilJ'ere ilt ()rsallisrlls. Iil' the use

.1i:,:rr.. \ 3.c jn. is prorluccd titroLrgit ciolting the vaccine gcne\ int() l:.tclterit-hia coli.

09鼈ラ

E^*rpl"r "f R".ombinant DNA systems Used to Modify Gene Expression

AcrinorhodinCellulaseRecombinant protein albumin

Heierologous protein

Enhanced gro\.. th i:il.".-\irlro aciri.:

St repirnnt t t s coeI itoIorClostridium genes in Bacrllus

S a c c h a ro t t t t c e s t- e re v i s i a e

Scccft arornr ce.i t' e ret i s i u e

,4.tpe rqi I ! u: nttlu litns(.',,t^,tt lu; t, t iiitti

Modification of gene lranscription

Arnplification of secretion through chromosomal DNA amplification

Fusiun tu a high-pruduction protein

Use of the inducible strong hybrid promoter UAS."1/CYCI

f)\'erproduction of glyceraideh-'-de-3-phosphate dehydrogenase

l:L)lxijL)r rri bio:r'nthetic Sene-\ rhat lead to enhanced enzlme :ctir ities or

r( , .Jr,'1 t:rJbr.k r.lLlJIl\\n

こゝ ostcttadrd:LO`sni mdJヽ た1%n2α"Mctab01で

cnsin∝ ing Or,ε「′口

"″)″∫εご″■5滋ξν:ε ,々夕ο′″ο′θiοι Ra■ 6“

'I)■50 Tablcl p 35

968

427 Choosing Microorganisms for lndustrial Microbioloqy and Biotechnology 969

ム

4ヽodule 2 Fvlodule 4

lヽodule l Modu e 3 lv4odule 5

Modified Structures

卜Иodule 2 4ヽodule 4 卜Иodule 6

Modu el 卜Иodule 3 4ヽodule 5X

3 ocked

einzy『 ln e

rv4odule 2 圧4odtile 4 卜4odule 6

Иヽodule l lt4odule 3 Avlodule 5

XBIockedenzyrne

，Ｓコ一】ｔ

Ｏ　ＯＳ聟ヽ「工一

Ｏ

C)thcl c、 tll1lPIC｀ illcltl(lc ti]c il]ビ rcti、 c(1、 ):]111c、 i卜 (,i tt1ltibiぃ tic、

al〕 d ccllulasc、 11,()(11lCtitiぃ 〕()1 ビc11ご じ` |)rCヽ i`t'I]l))ヽ へ al111っ lil'ictl―

11()13 grctltcr I)1()(cill )ヽ11[llC｀ 1｀ _ tin(1 11]1ご l tictiヽ C C11/)1llC ()ヽ Cllつ l｀ )(ltic

tioll o「 l c11〕 ●ヽ 111い 1 ltctibtit k i1lhilDit,ぃ :l Rcc● 1111｀ :Illlnl l)lょ 、1lli11ぃ ピc11

11)rc、 こ11111)lC 11｀ tl)(()111lD「 i｀ C ]()(o■ 1)`, ぃl thc 、、)l tl卜 Ic l)「 ぃicin ili ti

nlo(111lcd、 llaill a tc1litヽ 1(l iI]cl c(1ヽ C i11ビ ぃ1lCC1111を 11101〕 ()、 cl that ill lhc

oligint、 〕、tl‐ aill

Natural Genetic Engineering

Thc nc、vcst approtlch ilol crctlti1lg nc、 、 :1lCttib● lic capabilitics i11

a gi、′en ■licroorganisnl i、 tllc arc[l ol natural genetic engl‐

neering,、 vhich clnplo).、 frlrced e、 olution and adapti、 ie nlu‐

tations r、 どど/,″ 2∫ 9-■`', 1‐

his i、 lhc P!ぃ cc、、ol uヽ ing、 lDcc illic

cn、‐iI()innlcntal strc、 cヽヽ tい ｀「(ヽ rcc 1llicrぃ ぃ1■ tl i'iヽ nl、 1(, 111ヒ ItatC

tind をldtlPt thヒ 1、 cFCtitinピ 1111ご iて (`|「

=till i｀

子]]、 、ヽi〔 1l 1lc、、 hiい |ヽ iゞctli

CaPCthilitic、 1｀ hc 11lcピ Il till i、 In、 ●i tllビ ごヽ tidtiI)li｀ C nl tl〔 まiい ntll

P10CC｀ C`｀ incitldc Dヽ A「ぜtl lユ l11ご t i11● 1l t、 in、、hiピ ll irtin、1■

)｀ tilコ lC

clclllcnt、 and、 arious t卜 pc、 )`f rcct)1lRhirltllion Iつ i ll)critiCtil r()lc、

as、 ho、 n in tablc 42.5.

HO%′

一

.へ lthough thclc i、 llltlビ 11ご ()11[1て ヽヽCrヽ )CO1lCClllilng this arca,thc

lC｀ lD011｀ Cヽ いI 11]icl o()igtilli、 111、 1い 1ヽ1せ ヽヽ lD「

い`‐1(lc thc po(cntial()J

=CnCrtll l〕

g nlicrool=alli｀ 1ュ 〕、 、ヽith ncヽ 、 nlicr()bitil c1lpabilitics lk)「

tl、 c ill i1ltit卜 tl itl1 1]licI()IP i()1ぃ ぎ)tinti t)i()lcchl101()g).

Preservation of Microorganisms

(:)ncc ll nlicroo「 q(11l i、 11l ol 、l rtl、 1lt卜 bccn 、clcctcd or crc〔 itcd to

scr、 ca、 PcciliC purP()｀ C il intl、 t bc Prc、 cr、 cd in its original fornl

「or itll■ hcr usc and sttld、 Pcliodic tra11、 lcrs of cultllres havc bccn

t、 cd in thc ptl、 t、 altilougll thi、 can lcad(o lllutations and Phcno―

t),Pic changcs in lnlicroo「 gani、 1ln、 Tl)a、 oid thcsc Problcllls、 a va―

1lcI卜 ol culttlrc prc、 crヽ ation tcchnicltics lna)bC tiscd to lllaintain

tic、 ircdご じlttiTc chtirtictcli、 tic、 ltal〕 le 42.61 1´ }ophilization,or

r「cc∠ご_til卜 i11= tilkiヽ 1■ I tlビ ごiI' 11(ltliti llilliヽ

=cil ttiC trCqucntl).clll―

「lて いきti｀ i`tll 1llic:ぃ ●ritilliヽ i■ 、 へltht)ti=1l lぃ I`Dhiliztitiollを ln(l li(luid

ilitrて 、ごcl]｀ 1('I tlゞ C CtiC C● :い lDliCtit(:d tllnd rc_ltiil‐ CC、 Pc ll｀ 1｀ C Cquip―

nlcnt thc、 do tlHo、 、 11liC1 0bial ctilturc、 lo be storcd ft)r ycars

、ヽ1lh(ヽ Lll ll｀ ヽヽ ぃま｀、itl卜 ilit、 ()r til][lCCunlultitinil of nltltations

OO

O

lv・ odule 6

SS^- n:::--<".<O:.=

).,'rr HHo,.,. r> o, ---<HO> HO', /

', .<

" HO>i)

HO"

O

HO“

HOr,,

O

HO‖ "

HO HOJⅢ ‐

HO

一

DEB

970 Chapter 42 lndustrial Microbiology and Biotechnology 428

Natural Genetic Engineering Systems Bacteria

Genetic Engineering \{echanisms DNA Changes Mediated

Localized SOS mutagenesis

Adapted 1ia..nes hi iii n g

l-n-i.'1-n9. Tn.i 0 precise excistott

In vivo delction. inr ersion. lusion. anC duplicationftrrnration

Tll" II topoi.rn]cra5e l(cunihincll,,nSitc-specrl rc iecombinatittn (tvpc i trpoisomerases)

Trlrnrpo'ahle ricrtrents r1116r rpecir:r

Dl'NA uptake (transforrnation competence)

Base -substitutioirs. iranreshitis

i frameshil'tingRecipriral recon.ibinaiilr, oi'1lanking 8/9 bp rcpeais: ,e\iores orlgin,ll sequettc.

Cenera111, recip:ocll recombinlrlion of shorl sequei-.ce reireal\: occasionalll nonholttolrgou:,

Deletions anC fusions Lrv nonhomolotc)us iec()rllbinatjon. sometimes at shorl lepeu'rs

Inse(ions- ercisions/deletiitns. irr ersicrns bY concerterj or succcssir e cieaYage ligati(ln ieaciir)It\ Jt

\horl \(qllella. Ien. JI.: i,,icrll(. ;itt.;t.:tlchc.Insertigns, 11ansnositions. rcplicon 1lsi1ns. arijacert deietions/excisions. adiacent inr cisions br 1ig:iticin

of 3'Cll transp.r ton ends of 5' PO. grouirs llom silggered cuts at nonhonrulosou. tarscl .iLc.

Uptake o1'single stranrl indepcndrni ol se(lllcn.c. or of ri<;uble-slranded DNA carrvin.t spc, ie.identillei'sequence

Adap{ed from L A. Shapiro- 1 999. Naiu.al generic engin€ring. adaplive mutalion. and baclerial .volution. In MiL robial EL tios\ oI lnfedit)us D isedt?

for Microbjology. Derived from Table 2, pp.263-64.

E Rosenb,:rg. editor.159-75 washinglofl- D C: Americrn Sditlr

Methods Useci to Preserve Cultures of lnterest for lndustrial Microbiology and Biotechnology

■lletilod L'0mmerlts

Periodic transfer

N{ineral oiL slant

N,linimai metlirrm. clisiiIlerl qater. or'.\nter agar

Freezing in gros'th nredia

Dry uig

Frrrz--Jrr rrrl,.ropl:rirz;rl.,r, t

Ultrafieezing

Variables clperiodic transfer to new media include transfer liequency. medium used. and hoidingtemperature; this can lead to i ncreased mutation rales and production of i'ariants

A stock culture is grown on a slant and covered rvith ,rierilized mineral oil. the slant can be stored at

refri.gerator temperature

Washed cultures are stored under refrigeration: these cultures can be viable fbr 3 to 5 months or longer

Not reliable: can result in damage to microbial structures: *,ith some microorganisms. however. this can

be a ureful meanr ol cullure lnrinlenance

Cultures are dried on sterile soil (soil stocks). on sterjie filter paper disks. or in gelatin drops: these can be

stored in a desiccator a! retiigeration telrperature. or frozen to improve viability

Water is removed by sublimation. in the presence of a cryoprotectiVe agent: sealing in an arrpi:le can lead

to long-term vrability. u'ith 30 vears having been reported

Liquid nitrogen at I 96'C is used. and cultures of fastidious mrcroorganisms have been preserved ibrmore than 15 years

4.

1. What types of recombinant DNA technrques are being used to

modify gene expression in microorganisms?

Define metabolic control engineering, metabolic pathw/ay en-

gineering, forced evolution, and adaptive mutations.

Why might natural genetic engineering be useful in modern

mrcrobial biotechnology?

What approaches can be used for the preservation of microor-

ganisms?

MICR00RGANISM GROulTH INCONTROLLED ENV:RONMENTS

For nrani industrial pr'r)cesscs. inicroorgani\nrs rrru:t ht gr,.,"i n

using specificallr designed rredia undcr careftillr contlollcilcondiiions. including te mpelxturc. rerlti()n. and nutrient ieed-in-e. The developnrent of appropriate culture nteCia and tlle

!f()\,, 1h 1)1 rliar()()r:-iini.llr\ Lmder rrlrltrrtriril etlntiltiotls llrd the

rLrbicct' r)1' lil\ \aaii!rnI3.'iolr'1,'1 ,,.'1'q',lrnt. rl i\ naaai\:rr\ to eilrjtr lailltilloi(lg\. Thi

ict-nt ltrntttrtation. i:'i'ri tn lr Ph-r:iolLrgical \cn\L' ln arrrllcr-ra!ti\rrtl,rl i]lq l)1rirk. r\ anrIi(r\t'tl iir lt nruiir nrot't gettenrl ,,rar ttt rel:ttiott irt

intlLr.trrrrl nri.ir,broloSi linil hi()teahnolrigr. As noiutl itt table -12.7.

the tarnr alilr h:irt sercral nrr-rrning\. rneluding thc nrass ctllttrre (linricrorrrguni:nr\ (r)r c\en plartt alrd anintal cell\|. Thc derclopnlc-tlt

of industrial ternte nlation: rL'qLlil c\ uPprr )prl]lc uulture llle dia alld

the large \cale screening of nricrooreanisrl'rs. Otien r enrs are neede d

to achieve optillrunr product vields. il{anv isolates are tested lbrtheii abilitr t() \\'nthc\ilr u lre\\ prodLlcl in the iicsirecl qr,lantrt\'. Fcu

are succe<.iirl

Medium Development

Tite itie.liuill .iiiJi,.r--ji,i,\ -r iill!T(,,rr-g.irli.tll i.,IrLrr:il ilc-cllil:.- ltCltll

influence thc :iolrLrrrre crnrpetitir enes5 oi a p&liculi,u proaa\s. Fre'qucnil\. lo,,,,ct-a..1 anrLlr nlntcrillls are uscd i\ sOLlrces 0i carbon.

一蓼

Fermentation:

「

_

Microorganism Growth in Controlled Environments 9■

429

3

4

6

A hrVord with Manythe Microbio t

An-r prtice ss involvrng the mass culture of microorganisnls either

aerobic or anaerohic

1n,1' blological process that occuis

Food spoilage

source Ra,Materl11 _____―一 ― ― 一 一 一

~~~一― 一 ―

Ca:-bon and energv \'{olassesWhey

GrainsAcr icultu[a] wasies (corncobs) '

Corn-sieeP liqutlr

So),bean meal

Stick liqgor (siaughterhouse producrs)

Arntnonia and atln'qniurt salts

l- itratesDistilier's soiubles

Crude preparatitlns oi plant and aninlal protiucts

Crude ino:qi11llc chcnicals

Chirlk or crurie cal blrtlale;

Fcrtii r zer-graile PhosPhates

Hightr alcohols

SiiiconesNaturill e \tal s

LlLrcl artd r tgetahie oils

ヽitrogen

irr the abscilcc of O.

fhc production of alcchollc bcvel ages

Use of an organic subsuare as the electrr:n doror and a'cePtoi

L:e of an organic -\ubstl ete as a reductanl and of the saine partiaill

degaded orgallic substrate as an oriilalt

Gr,r.tth clependent on substfate-level phosphorylation

,1い ｀`ll｀ t()P10(ltlCtio11 01に lCSil‐Cd lllctabOlitcヽ

Grovヽ√th Of MiCroorganlsttSin arl lndustrial Setting

l'he O:.()ileelltliliioll lrn(1 1'lLl\ rillt llltl\! be 'lll'li'iclltll illgh tir

hl, e (l- ill .:rccss \\ ilhill lhe cells so li'rrit ii i: not 1;nritirlg. Tili: ls

arsllccilill\ ltl.tc $ ht'll ll de llse iuic|ilhjlrl 'illturd 't gl'1111 111g' \\'hctt

l'ilarncntttrrs i'r-ingi and iictillolll\ cctc\ lila itlltLllcille ration cilrl ilt-

c\r-n lul'thcr llrliitcd bl l'illrrllentoLls sr\r\\th ttigure'12'-ll' Such

filarle ntotts gtou th fe stlils itl a viscr'ti: 111rL:tie rlle r'liLlill' kno\\ 11

as 11 non-Ne$ttlnian broth' \\ilich o1l'ei\ d\cll IrrllL'IL-\i\i'1ll!( ir)

sti rrittg llll(l ilerllli()ll.It is esscntial t() lisstlre that Lht':e pilr 'lclil 1'lr''1or' lrlt- llot litll

i,i,," ,11!('rohiltl sron th. 'f his i\ tlt"'l ''riticril tirrllrll scaleup'

there a sLlcce ssfill procedLlr- de reloptcl rtl li strr'ill shakc lllrsk is

nrodificr-l lor use in a iaiqe 1'erttletltet ()t.ie lltLtsi [lnder\tiil]d the

microell\'ilolltlletrt tl1'thc'cultLire lttld tllliitltaiti 'intilet ctlllilitiotl'

neartheindividualcclldi:spitcitlcr"lise:inth':tltlturcrirlttt-ttt'I1'a sttcccssful irailsititln cllll b' tllade it()lli 11 fr()'L-\s ()riglnitll\

Figure 42'4 Filanrentous (irosth During

Fe-rmentation. Filatl'itltlttLr' ltrngr rrntl sctinolll\ celc\

can change thsir glouih tornl tlttring tile course ola ier-

rnentalio;. The clcrelopr.nent ol pelleted grouth trr

fungi has nlajor L'tl'ects on ()\\'gen transfcr ancl energ\

,"qii,",l to a!itate the culture' (a t lnitial culturc (b) \l-

ter l8 houls sro\\ th'

Vitanlin.,c'Ir()n. trace siilts

B uiiers

Antifbani ltgents

fvtrj* Co*Ponents of GrowthUsed in lndustrial Ptglgsses

けChapter 42 lndustrial Microbiology and Biotechnology

Figure 42.5 lndustrial Stirred Fe rnienlers. ra ) Liu::r 1ci'

illarllal' u.e,l b) ii ])ll.rr'nlrLtIir.a] it,ntl.:l|ti 1irItltt nticr0l.rl] itrr,-ducliOrr Ul',rnliltioirt.. rltr I)eitriir rr1 il 1L-rntL.nlat Ltnit.'i-lri. Lrnit

cen l.c nln urirlci lL'r'!)hia ,)L ;ilt.tL'r'()LlL conclitiort:. rtnd itutiidnlittltltit0tl:.:arrrltlrrg..rllrl tr-nIrlt1.t{ion nt0,,it0r-in! eltrt l.c caIr'rcdoul Ll|(la| 11\al)t rt e\rrldil j()ns. Il i()\rn\{)r-s antl inl l -tl crr nli rl| [{ rt-r ilreatt ttrttridc riitl linrt inlolntltlion ()lt thc c()uf\e rtl lhc ltrnterrlli-tiolr. Speril-i. :ubstrllts. rrrelelrolic iltlcrrnediate:. alttl linal pr-r,d-

LlCt\ rlill b.' rlr'tcclctl

tle rt'loptd in rr l-i{j ntl I--rlcnilcrcr lllisk tL, rr l0( 1.()0() litcr r::ii,tor. thr'n thc t-rrof C:r i)1 :alilaUJr hli. t.r,r,lt I iill-]erl ,rul f ritIe rlr.

\1re lrrLrulinisrn\ Li:it ire qto,.,, n r:t crrllurc lrrhcs. shakr. 1'llisks.

Itncl :liilccl 1c't'ttttnlcr'' ()i otitaj illlr\\ LLl llLlta \\ \Lcnt\. Stirrctl lirrllL'iiL.l\ eiln [l]l!e iir 'rze llont.l LI J li1.'r: to liX).000 Iitr-rs rtr'

llu'rei. rlcpcndirrL on lr'rrductjr)11 trrlruiL'ntclrl\ {{igure -12.-it ,\tr1ri1;11 indu:trial \1nrdd i'arntcnliitl()n unit i. illLi:trutcil in f igLrrcil 5/,. \r,t,url., ilL:st 1]1. nterliiillt he:l.,rillu.-tl lirLtt tar'.i11()n. i)HJcl.iir.lnteni. .:ilI1t1i n.l. liltil ltr rtc,:.s rtl0nilirIttlg lltLt\i bc cet-r-i,:il

r jLlt LrnrlL'r ri-rolouslr clntrolletl eonrliltorii. \\ hert rrqLrired. ioaltL()ntrol lrgcnt.\ lIr.r\t he lrtided. r.\JleciJll\ rr ith lrigh Itroteirt ntr'dia.Cttrttptrters arc conlttonl\ u-(ed to ntonitor outltut( f ilrn probesthut deter ritine lliu r(rhiel bio1111155. lr-r e is ol critical ntetah()licprod,lcts. p.H. inpul antj crhaLrst gus contposition. and other pa,ranrelers. Environntenllil conditittns can Lrc changed or hcld constant over time. de penclin!. on the -goals f()r the particLtlar proces:r.

Frequ.-ntlv a e I il jatl e ()litp()ltent in ihc titctliunt. olicn the clir-boir slrrree. i< li.i,l..il a()niinu()Li\l\ iontinuous feed.-so thatiht liiiarLrrril:ilti:nt ii tll ::,,i i:-rr i. i\!tL'.\ .;-th.ii.tii lrr:illlrhle ltl anrli',r-rl iittlt l-hi. i. !:1iir!ri,:1 lr 1;11|, ,:[.i',] .,t'ir Iil.,r:r- iind Othl-rJilrhlrltldriilai. II a\1i.. Liriao\a i\ J,.r'a\anl .ti iht br-sinltirr:: ()f a

1iL'ilrcntrltron. it can be ratabLrliz.-il to r ie ld e ihlnol. rihich is lost

Culiure ornutrient aciditicn Dissolved oxygen prol

Coolingwaterout

Air in

lu: l rolrrtiic lrotlLrr't uitrl r-etiLrces thc f ilril lielil. Thii clln ()cc1

i'\ rn rrndei ltc'r'oirit c()nriition-\.Ilesiclc: llte traciitionui stirr-erl aer't)hic ()r unal-r()bic f'enrenle

othcl lrpprolrchcs cirn Lr.' uscrl to gr(i\\ rrricli,rrganisnts. These a

ternlti.,cs. illristrrrtetl rn figure 42.6. includc irtt-tube i.nlenlertligirlr' 11.(rrr). uliiih clintinrite thc neecl li,r:tirrcrs that can L:

loLtled h,r'f illutcntoLts 1Ingi. Also arailahlc. js:rrlrr-1-state ferrnertrrtjon Iirgulc -+1.(r/r. irr * hich li plirticulatc \ubstrate rs keltt ntoirto nlrrntliin;t tlrin sLrrl'ltce riater i'ilnt uhcrc nticrtibes ciln gr()1

rnrl orvgen is aiaillrblc. In rarious l1'pes oli'i\ed, (l'icure.+2.(rc

and t-luidrzed-bed reactors (figure 42.6d)- the micrtrurulnj'nts lrassirciated u.ith ine'r-t surfaces as bittfilrns (see pp. 6(1.?--l). an

nredlurrj f-lou s past the fixed or suspended particles.Dialysis culture units also can be used (figure 42.6c.t. Thes

units allor.r toric wliste metabolites or end products to ditfusau av fl'om the microbial culture and permit neu,substrates to cliffuse thrtru-rrh the nrenrbrane tou,ard the culture. Continuous cultLrrc tr-chniqLres using chemostltts (figure.l2.6l) can ntarkcdlinrpror e ce 11 ouiprits liltri rttc\ ()1'srrhitrate uSt br-Cl1u-sc ntict-r)\)r!iini.ln\ can he ntlintained in a continut)i,ls iogtrithnlie phase

1-{ou cr e r. continLi(lrri ntaintenancc ol an ot.gani\nl in en litir,gro\\th phase is undesirable in rnanv industrial processes.

42.

ｅ

一鏑‐ｉｎｅ

972

pH probe

二

・ 42.11

{a) Lift-tube lermenter

Density difference of

gas bubbles entrained

in medium results in

flurd circulation

Microorganism Growth in Controlled Environments 973

(b) Solid-state fermentation

Growth of culturewithout Presence of

added free ',ater

(c) Fixed-bed reactor

l,4icroorganisms on surlaces

of suPPod material:

flow can be uP or down

(d) FluiCized-bed reactor

lv'1!ciocrganisn-ls cl sur{aces

o{ particles susPenCecl

in liquid or Eas strean '-upwarC:iovr

FlovJin+:

―Ar n

FloヽV Out

F orv out

SuspendedsupPorl Padicles

FoW n 一

(e) DialYsis culture unit

Waste Products ilfluseaway from ihe culture'

Subslrate may ciifiuse

through membran€ to

the cuiture

(0 Continuous culture unit (Chemostat)

Medium in and excess

mediLrm tc v/aste \Yittr

vJasted celis

Figure 42.6 -{ltcrnate \lethods ftrr }lass

(.JIture. ln atlclitiorr to stirrctl fr:rnlcniers' othet

lrr(llt',r1. . .ttt irc tt'."1 i(r L rlllrllt' llli' i""t j'tlll'ltlt ill

urr-ltt:tnril ilr()cas\ci' lti Inliilr 'ases

tilLlse altel'nale

lrnlll olichc' \\ il1 ha\ c 1(r\\ er ()lleratlllg costs alld. car

1,,.,.,.t. .irc'illiztJ !l ()\\ th'()ilditi(rll\ tlcecletl i'tlt

Ir rrriLrcl .\ 1lt1-la\i\

1ビ lalion、 hip to lhCヽ 'ゝ1lthCsiヽ ()「 cCll 11latcrial、 alld l10r,1lal groヽ 'th

lヽoヽ t antibioticヽ anci thC lllゝ COtO｀ ins fall intO this catC80ry

t. Uo* is the cost of media reduced during industrial operations?

;;;r;t iire effect of changing balances in.nutrients such as

tr4edium rn-*€

\ Iicr-ohi;i1 Pltiduct\ tlltcn arc cla::ifit-c1 as prilllllr\ and :ec'

ondar\ metabr)lite s. As si.torvn in figure t12.7. prinrar] metabo-

lites consist of cotripouncls reirtcd to the s\nthesi-i of migrobial

celis in thc grou'lh phase Thel inciutle ainirlo.ilcids' nucieotides'

and f.rrnentation entl orc"it'cts stlch 3s ethlirlol rrnd organic acids'

In adLiition. industriailr us'1'Ll1 L-nz\ 1l)!'\' e ithcr rrsttlciatecl u ilh

1|1g 11i;r1rl1ial scil: trr i\t'c1l/)111r" \'1t'll '11'- \\ilthc\iz'i fr iltr-

cr(rttt !:ttli:nlr,1url ilS !rL'ri tfi'

Secondarl metab()lites u\Li;illi ii!''tirlitllJi' jLititl: thr i-t-

rir-rrl ot nutriL-ttt litl'titatlon t)J \\ lirie pttlrii'tcl 'tiii:lll''illtitltl that iLrl-

lo$s the acti\c sro\\th phrtc The:e ctrlllplrllnds hare no ditect

minerals′ 910ヽγth factors′ and the SOtl「 CeS Of carbon′ nitrOgen′

and Phosohorus2\.^VhatarenCn-NewtonianbrothsandWhyaretheseimporlant

in fermentatrons?

3'DiscussSCaleupandtheobjeciiveofthescaleupprocess'

「

chapter 42 1ndustrialヽ 4i(robiology and BiOteChnology

5. Besides the aerated, siirred fermenter, what other alternatives

areavaiiabieforthemasscultureofmicroorganismsinindus.trial processes? What is ihe principle by which a ciialysis culture

What parameters can be monitored in a modern' large-scale

industrial fermentation?

system iunctions?

MAJOR PRODUσ S OFINDUSTR:ALMiCR08:OLOGY

Time

pre 42.7 Prinrarr anrl St'c<tridarl \letahrllites' I)e-

r,1ii,g ,,n tiic llalricLrilrr-otrllllll\llr' ihc tlt:ii'ril llrorlltet tlllti bc

,ra.al .1r.,ring or a1'ter o""tth Pi 'iirlrr tllttltboltlts lllLl 1(rlllle(i

-ing thc ,,ati, e gro'n'th ph:l-'c' u ile tclts st'c(rll(l'lr) lile 1'tbLrlite s

forrttetl atier grou th is cirllllllctctl'

Iniiustrilrl nrlctc,L.iolcigr hlts Ptt'rr idt'd prodLlel's that haYe in pro-

1'oLrrtCli' cilirngcd iir-rr li" es :rnri iiie -tpans The y include industrial

lnd ,.rglictrlrLrrlil prrtduclt' 1'ood lrdditir es' iliedical products lor

lrun,,,l unci rinjllii 1-rtalth. rtnd i-'ioir-rels itahle'12'9i Particularly"

in rhe iasl 1e',i r erirs- llonillliibi()ric !olllp\)rtlld-\ rtsed in metiicine

ancl healtit hure tliltrlc tilrijLit c,,tlitihLltiorls ttr the itnprrlved u'ell-

beinc o1'eililllal lrll,l hilitlrln polluf iiitlns' Onlr l-tllijor products in

r-i-re h arite!(rl-\ u ill lle tli'' ti'<cti ilr-rC'

Antibiotics

\lanr lrttiltrrrlla .lla 1)t'rtltltr'ti hi tllltl(r{rl!Jllislll\' preclonli-

ii1i1-,i]r lri iiiiiil(rill\ltl.ls :ll tht geitLts !/r1'r711r17;-'tt'' lrld b)'fila-

,,,a,,,,,u.'fLItlgt i.ii'. tirhlr' -i-i 'l' irl "l

' ; ehlllter' thc irnthesis of

seretlll i)1 tlrt'Ill(l\1 lllll'1i)llrrlt ltrltibitttl-: rrill be tiiscusscd to il-

ndustrial I'roductsEthanol (froin gluco-se i

Ethanol r:from l:lctose )

Acetone and butanol2.3-butanediLrlEnzl'mes

rgricultural ProducLsGibberellin-s

'ood AdditiresAmino acids (t.g . 1r sine t

Or-canic acids (citric aci'ilNucleotides\4taminsPolysaccharides

vledical ProductsAntibiot'icsAlkaloidsSteroid rran.'rfonnations

lnsulin. hunran grou th htlnlone' som3tosi'iir

BiofuelsH1'drogenN'lethaneE:thanol

-Sur'aftrrltriltr at': t {'r?1 1Jl(?('

K Itttt c rontt r t s .fra gilis

Clo st rid t Ltrrt ttce I ctbutt I i ttrtrt

Ent e t ol.tatt e r, St rrai ia

.{,t1rcrgi11iir, Bttt.iIIus l\1tt{ ir' 7 tt'-lndIrn1Lt

Ciltbtn'![o.fujLkunt

C o n ne btttt t' ri tt n 3l ul arn i t t tttAspergiLlus nlgr:t

C or : ne bat'le ri tt ttt 3l ut tt rtt i c tttrt

,\ :hb: tt. E re nu nlrc r'i ttnt B I ttkt sl t' tt

Xtutllrcntotns

Peni t' ill ium. St rep lontt.<:e s' Bac i I lt t's

ClavicePs PuryureuRlrr:,,r,u' 1'111"'l'a' 17r

Est Jier ichia t't',Ii \titcJruronu "s "/1'l lsl(l''

.;-., ith'r:,:' tr\A 'e' l-n 1 '':i

Phuir:r nlhctii miir,'itr:slisl:',\1 e r I i tt nt t b ti c ! L t i u nt

Zt'momon as. T he rnn onae ro bacl e t

and oth:i-s1「ltCl■ CrOnヽ

:' Prirnary- metabolite

な）

formation

Growlh ,/

、、

ヽ ｀有

; .." Secondary- metabolite

formation

Major Products of lndustrial Microbiology4213

（ＴＯｒｘ

ｏヽ〓くの）Ｃ〓〓ＯＦ一ｏ０

ぃ０ア』ｏＦＦＥｏ

０“０」０ゝ〓００」ＯＯ

．でｏ〓ヽご）のりｏＦ一〇一ｍ

o ?o 4A 60 80 100 12n 140

Fermentation time (hours)

Figure 42.8 Penicillin Ferlrtentation Iltrolres Ilrtcisc(ltlntrol of Nutrients. 'l-hc s)'rltllcsis o1'Perricillin hrgills n lt"rl

rrittt,-le tt 1't'titlt ltlttttlilt-tia becoltler lirnit;ng' '\1-tcr ltltltt ot'tht- lltc

t6s.. rlr sipl 1r cutahplizcd ilislrer'hlrt iclc)llri: fictlt dcgllded. r1ir

cr)sLt (it rlrpidll Ltsctl tlltlllrlslicchliritle) i: liddctl :lloll-l rr lth 'i lorr

icr el ,.rl- nitritge n_ Th js srintLtlate s nL[.inrunt tt litlsl'trrtrllititllr r)lthr'

cai-ltLrlt \otlrccs ttl llclliciliirl. Illt scllit t'alt()l i\ Irl tst'lrlttl tIitl-

the crtlt\etttii)n fec()illlllrrlldctl br t lle'\SNl"iilat l5' 1 11li111[1'r. t"t'i

the aris shoitld be IlrLrltiplii:d br'0.10 to olrtliirl tilc trttr'ralLte '

lustl‐ titc thC Clitical role of incdiuinn forlllultlti()n and cnvil‐olllncn―

tal contl‐ ol in thc productio13 of thcsc inlpoltant colllpounds 蠣ヽ

tll),i ti、 1 ,t、 、lit t lt■ 1ヽビ: 「` |

Penicillin

恥ぶ[1鮮議I勲熱、11#:Ⅱf鷺

i研硝‖篤胤烹博 諜∬だ11ゴ賓I葛Iぶ

‖窯∬)点‖I鷲∬fttIIT∬‖鷲Fζ‖ボヽ性R満樵mtibiolic icヽ ctt inぬ c culm「c hcginよ iヽn劇で■、ctngurC 42.9,un―dcl ct,11こ liti(、 11ヽ いic● 111「011Cd 1litlo=Clユ li1l111[ttiol〕

Anlino Acids

へ1ュ 〕i1lo[lC iti｀ tヽlch ti、 1卜 、111ビ [IR(1ピ 1ヒ ltユ 11lic tlご iti ttせ ょヽCd in tllc lkヽ いに1

ilndtl、 tlヽ aヽ 1l tlt■ tl● I〕 tilヽ upplclllcnts in blctld Prて tヽltiCiS tlind tl、 1lti、 or―

cnhancin=ご 、)11lPい und、 、uch t卜 1110n()ヽ odiunlピ lutanlatc t｀ lSGI

Figure 42.9 Streptonrlcin Production br

5tr-tpt,tmrctsgri-serls. L)tIlci"''- "i jiii''i':' lelii: 1o

nriL\iillunl antihiotic I icids.

０●一●

，

〓

。

３

Ｅ

ＯせヽっＥ

】

Ｅ

５〓ｏＯか

，一つＥＯ

（，

こヽゅ〓し

ｏＯｏ●ョ一ぃ

いＯ

ＣＯ〓●Ｌ”‘ｏ●●●０

奮Ｆ一ヽい、）Ｃｏ■ｏし″Ｃ●ＯＣｏ●

●一０、Ｅ

Ｏ″αｏ」ヤ∽

975

― ―

Chapter 42 lndustrial Microbiology and Biotechnology

ｔｅ

　

　

　

　

　

　

　

　

ａｔ

ｔａ

ヽ

１

１

１

‐

―

ァ

‐ｔｒ

＼、、、、　　　　　　軒『一′

a)

le hv Coi-r'rebntterirtt'rt qlulamicum. \ 1a.jor cer.hon tlrtws arc notcd by bold arrows(a)Growth with usc ofthc gi)()、 卜late bypass to

shitjccl holtj r|.ro$s) 'l he dlisiterl line-s ind;cate rL'lcti()ns ti-litt lre bcrllg used to a lesset e\tenl.

42.14

ｓｅ　　　　　ｏｓ

∞―十

ｐｈＧ‐ｕ　　　／／／／／一

ａｔ

　

　

　

　

　

　

　

　

　

　

　

Ａ

ｐｈ

ｌ

ｌ

十

ｃ

T百 ose pho

|CO

/ > Acetyl-coAC

CO

封r

)γ

」:te lileAcetyl-CoA

CHOC00~

Glyoxyl21s,,

′Oxa osucc nate

r

●Kao― ″・島

91utarate ‐

INH+

T

G utalnate ti))

Oxaiosuccinale4.

,,-n",o @*& UU.

g NH.**w

Glutamate

＼ヽ

Fumarate

I\\lr

Succinate

\ \Suce 1nYl-CcA

I

lsocitrate lyase

cis-Aconitate

\I

I- - - lsocitrate

&6sF

.!Irn() :iai11 pt'orittctton is t1'picall-r' cal-ric(i ()tlt b\ lllcllrl\ { )1

latorr ntutatrts. u lricit itavc lt redLlct'd ahilitl to liitlit svn-

. rr1 liir r'ttLi pir)rlLlal .f

h.' ltrlrnlal illicrool-lliillisrl ltr oitls tl!e r

l.ti(rn ()1'l-ri,rlhe lrillrl inlr:rnreriiltes b,r'the carefirl lcgulllli(lllllulur rlcirih,rli-ittl l'roduclion Lrf giutarllic ae id and scr eral

rllrnr)i1ai1i\i\Il()\\ al1l IiaLl outLlsrllgllltltxntsof ('rlr-t'llrl1.;ttt-

t: <lirtLrnitLirur tlllt leck. ilt htirc onl\'ll linlited ahilitr tcr

'cs. ti're lL \ crule IIttct-ltlediiite u ketog]Llla|Jle (.\ce dpPettr) t(-r \Ll.r'ln\ l-(lo-.\ as sh,.rwt.i it.i figure ;12.10. A ctlntrolled

riotin lerel uird the eddition oftattY acid derivatives results

:reasctl menlbrane permeabilit) and excretion of high con

:itions oi'siutamic acid. The impaired bacteria use the gl1'-

iie path$a)' (see set:lion 10.6) to meet their needs for

tial trrochen'rical interrnediate s. especially during the grou'th

'..A1ter gronth hecotle s lirnited because olchan-ged nutrient

:hilitr. rin lllnosl cott.tPletc nlolar conversiot-t (or 8I .79

tl a()n\er\l(ri) oi i\oaiirait t(l !luianlate occLlrs.

\ \iila. ilir a\scntiiil :lllllll(r llCltl usr-d 'ra) \ullPlerllent alefeal-\

rrc.1d\. '\ :is oriSinall\ ilrodu.e d in 3 t\\ o-step nlicrobial

PrOCCSS Tll:、 ha、 bccn rcplaccd b)を 1、 i llぎ Ic― tヽCp it「 IRlcnttltion in

、、hich t1lc l)actcritllll(｀ て,′l、′7こ′わで

`(rピ′′ι″77 .、 ル

`た

′′/111′ ″だ い1● ckcd iln

thc s、 1lthc、 i、 ol:honlo、 crillc ttCCtlllltlltltc、 1)ヽ lI〕 c ()ヽ cl■1=llitCI

Ctll]bC prOdtlccd iin t1 3(la、:fcrlllCnttltioll

Organi〔 Acids

(Drganic acid pl‐ odtiction by 11licro● Igtini、 l〕 1、 1、 111lPい lttillt ill i11-

dtlstrial 1lnicrobi(Dlogy and iHustratc、 thC cllicct、 ぃli ti tiCC I:3Cttll lc、 ―

cis and balanccs oln organic acid、 )ntlncsi、 and c、 crctiぃ Ii Citlic

acctic,lactic、 fuinaric,alld gluconic aci(ls arc nl轟 or prOducts

(table 42.10)Until nlicrobial processcsヽ crC dC｀ cloPcd tllC ina―

jor sourcc ofcitric acid was citrus tuit fronl ltalゝ 'Toda).most cit

ric acid is produccd by microorganisnls:70 r iヽ ヒl、 cd in thc food

and bcveragc indust]、 201を in Pharnlacculical、 and thc balancc

in othcr i1ldustrial applications

Thc csscncc of citlic acid fcrnlcnttitioi〕 i11、 |,1ヽ じ、li:lliti:1ビ thc

a11lounts of tracc nlctals such[1ヽ 111■ 11=tlll C｀ C t11こ l ir● 1l tt,、 tい

「 1ヽ―

/'ど′メ′/′″

'″′gど ′groヽ 'th at a sPccilliC lっ olllt ill thc icr,1lc1ltation l‐ hc

ゴヽ

'( > Aceiyl-CoA

CO:ツЮacdae / 6tde

Major Pr-oducts of lndustrial Microbiology 9774215

fVtrpt Orgrnic Acids Produced by Microbial Processes

Fermentation Conditions

Acetic iicid

Citric r.rd

Fumaric acid

Giuconrc acid

Itaconie lciti

Ko-jlc acid

Lactic acid

ism Used

Ac e t ob(1( 1 e r s'ith elhanol solutions

,4 sper.qi,'ltr.r rt i-ger i r: l.toias-ses brtsed

nredium

Rhi:opLts ttigrir:ans in sugar based

mcdiuin

Aspe r';illus /i!'gdr in glucose-inintrai

salts mediunt

..\;ptrqillu: lt'rrctts in mol15se\ salts

me d;un-i

,4 s pt rg i I I u s .fl tt rtrs oir'-rre incarbohvdr:rte tntlrgrnic N mediillll

ilomolirmentati !e a,i.1r,(l.tllll t

,it'l brur: t'kii

tatiYe Uses

Yr'ide variet-v oi' iood Llses

Pharnaceulictlis. as a lirod itdditir t'

Resin man,:lacture. talli'ling.:'rnrl sizing

A carrier tbr calcium and siidiurn

Esters can be Prtll''rterizedto make plastlcs

The manufacture ol fungicidesand insecticides u'i.ien crltllplexedu ilh melels

.\s a . rn-ier tor calciunl and

as an acidifier

Single-step ttxidation. *ith l5%solutions produced: 95-99% yields

I{igh carbohl'dr:lie concentrations and

cuntroiied lin.iitation of tr:ice metals:

60 80% vields

Strongll aerobic tcrmentation:carbon-nitrogen ratio is criticll: zinc

shoujd be limited: 60% f ields

Uses a-eitation or stirred fer:llenters:

9-i7r yields

I-lighli aerobic medium. below pl{ 2'2

35')i. vieldslron must be carefully controlled to

arojd rerction tirh kojic acid atier

i ernrcntationPurif iti mediunl used rrl tliilitate

eltracl iL)ll

Irrr-dirinr rtitctl i: treltlr'd utth totl d\uh3ll!e l.'tiil\ i!) 'll'tllc

l()\"

;jn(1 L()ltll!)llarl roitcr:lltflllir)11S ()i'lL\illlilllle tllCtlil'. illl!l l\ aillli'ri()ut in iiet.(lbia .tit rcd ielnt..nlL.t \. clerlL-rltilr. ili gtl \'l!iil L()ll! rrll

Ll.rii.'r.,ilr : r'r' rr'r'1..1.i,,'1'1,r11f i"ti i'i: i:' \''riri

Ierlct tllL i]lhii-tti,rrr oi eitrii li.id lll)tiLlctlon 1), il.t)il lrb.rt'().1

irprll. I hc stlrLt\\ Lrl thts lcrllltlltiitioll tie Ilt'nli: otl tht ltgtllltti()n

rrnci lirncti0ning ()1 titc ulr c()l\tic pilih\\ l\ lltlti 1ll.- Itie ltl ilirr) lit'

ilai(l !\cle 1ir.r' \{'(lit)tt !i.+)..Al'tcl tlle lLciirL'gl()\\1ll l)hli\'' \\lltll

1113 :11ir:1t.rrtc ler tl i' lirril. ciLI'ate \\ nthlst' ilLti\ it\ iilclclt\L'\ lirlrl

tltr- lriti" i11L'. {)t l.i(i)llliil\c illlLl i:0cilretc tlchrdro!e lllLsc dcrr':lL:c.-[]11'

1-sittlts irr iitiic ilaiLl ilucLilllLlllititln arlrl c\'1r-lioi'l hr the

sllL s.t'l lt,'' I ' :-.llri'll'.In !,)rtti.r:ii ir,rtl. iilt' llirrdLttlltrtl ()1'-Jlll'i)llic ilci(l i11\ rrl" l: lt 'itr

glc tlllr:rtrhilti cilzr lllr'. glLlcot3 o\idilsc' ilrLrnti tn l t/ri r'(tliii'

niqt'i: .1. rilqri l: !l()\1.11 Ltniltr (llltilltlllll coll(li1i(rll\ itl r '"111-.iealt jt(lu(rr ntctlir.till (rrtlirth bectltlles lirllrtctl br tlitl-i'!elt lintl

ihe rt-stiilg aall\ illlll\lorlll tiic lclllaillinS glLteose t'r !lLtcrrtlii 'tciil

rrl lt iril-!lt-slal) lr'11a11(r11. (ilLrcrltlic eciLl i: irsctl ii\;l (rilrri'l l'01 clrl-

aillill rlllrl i1(ril iiil(1 t( il Li)lll|()llr-llt 1r1 (ict'lgell1s'

Specialty Compounds for Usein Medicine and Health

ln atldition t() the trlllk pl odLlct\ that hare ilcrn prritluced tircr

the lasr -l()ro '1() \'cars. such a\ antibiotics' arllino acid\' and o1'-

ganic acids. illicroofganisljls alre u:ied ior the orodr'tctioll cli'

nonantibiotic specialt)' conlpounds. These include scx hor-

nlrlnL's. linlittllllor sgents. ir,noLthores. antl spcciltl '(llllp()ulld\

ihlt lnllr.icnce bactciia. tunri. atlrrehee' in:!-ct) iind fllllts,table-t2.l1r lrr lrllLlt:c). ll i\ llL-!c'\t':l:' t' ir"dilil rrittl Iitrr\il'ihe r,t-,,ttLicl. itirdci iaicl'rilli .irn1ri,11ct1 cirildlli'rir' l1r:i\\r'lij illlll

'Lht:t;r-lr-rilia]lr iillPt'rie;ll a(lll'lll(rtiIlLjr fL'llth illc'-rrll'!; illJT ril 'l

rrihlt. r1ltr l'\ i r "l'lJlli'rll.

1 What critical Iimiting factors are used 1n the penicillin and

strePtorni'ti n fermentations?

What are reguiatory mut2nts and how were they used to in-

crease the production of glutarnic acidby Clrynebacteilurn?

What is the principal ltmitation ci'eated io stimulate citric acid

accumuiatron by Aspergillus nrger?

4 Give some important specialty compounds that are produced

by the use of microorganisms

Biopolyiners

Biopol\'mers ltt'e tllicroi-rilill-r' llrodr'rced p()1\ illeri' prilllaril\i

1.,,lrsli.:ch,iri,l.s. il\ccl 1ri gttrdii) the ilo',r chtrlctcristics oi iiil-irirlr utt.l t() scl\c Lls ccllill! llgcilts. Thc-se llfe eniplr'r\'ed tn ll-tatlv

.rrcls o1 thc phar tllltcctlticll ilild lootl jndLrstrie: .' I

.\1lelr\lli'ioiallpoivsacchltt-ldesarcuscdirsstlihilizers'forilrc .ii:pr'r:totl ()i lrilrliltllllte-s- lrs f ilrrl-iorllling agenl\' or 10 pro-

ir(rlc ',i lte t t e tetltiilll ill I at iou: products. Ptrlr''lccharicle s he11l

ntiiintuin thc te \turc ol nlitn\ 1'rozcn lixlds. stlch ils iCe Creitlu. that

1i1.1 511f-iect to tlrasiic telllperatLlre challges' These polr:saccha|rdes

musl rraintairl their properties urtcler the pH conditierns in the par-

ricLrlar lbod and be compatible \\'ith other polysaccharides' The)'

sl-rould not losc their phy'sical characteristics if heated'

Biopolvmers also include (l) dextrans' which are used as

blotrci .jrpalrders atltl ahsol'bents: (2) Entinia polysaccharides

rhat lr'. us.d in plinis: anci (l) polvesters' derived fronl

F)\(,tLlttni()nti.r rrlr'r,r'rr'au1.r, nhich are a feedstock for specialtl

p1;1s11;r. L'tllu1L,.t'tlrtci(r1-illril\. plodLlcdd bt'trt '1t'ctoboc'tcr

.illin. rr. it\..1 r\ ll 1'r,,id thicke rle r. Polvsaccharides such as scle-

i{r!lLtcllt ale trsed b] rhe oii intlirstrr as drilling nlud additivcs'

t,

97e Chapter 42 lndustrial Microbiology and Biotechnology 42.16

Nonantibiotic Specialty Compounds Produced by Microorganisms

Pol),ethers

AvermectinsStari ns

Enzyme tnhibitors

BioherhicidesI;nnr u nos uppressants

Anabolic asents

UterocontraclantsAntitunrdr agents

S I re pt o mr c e.s c i ru ru t tn n e tt.s i s

S. losalien.sis.\. albus

S. averntitili.sAspereillu: krreusPet, ; c1 1 1 1,r,, c it rin unt I

aclinomycete'

S. clavaligeru.sAc'linoplanes sp.

S. hrgroscopit.u.s

To Irp oc I adi unt i rtJ ! a t tun

.5. tsuknbaensi.;

S. ltt e ru.; t r4t i t' L t.s'

Gibberclla :roeClaviceps puqtureoS. pertt:et ic u t suhsp. rrir:.irrri,5. lteucetiL.r.:.5'. i rzr'.tpilo-trr.s

N{onen si n

Lasaioc id

Salinom,vcin

LoYastatin

PraVxstaii.

Clavulanic acid

Acerho\i:

B iaiaphos

Cy,closporin AFK-506Rapaml'cirZearaienone

ErgLrt alkaioitl:Dororubi c i n

Dru noruh i ei n

\llrrrJl\arll

flleornr cin

Coccidiostat. rumenal grorvth prornoterCoccidiostat. runinal grori,th pit)n.toter

Coccidiostat. runtinal grorvth prornoterHelrninths and ailhropodsChoicsterol -lorverin-g lgen t

Cholesterol iou,erine agent

Penicillina.se inhibitor,Intestinal giucosidase inhiLriior klccre:ses h,r peigly cernra and

irislvccride s]'nthcsi-(l

Organ tr:rnsplanrs

Orean transp)ant:Organ transpllrnts

Farnr anintal medicaii0rInduction of laborL anaar treeimcntC:ncer treatilenl( itnaei trcrtlntnlL ancr'r lr i'lttntcnt

Compound Tlpe Source Specific Product Process/Organism Affected

"Compac(in. produced by Peniciilium citrinurz, is ch.nged to prr,.asElin bv iln aclinomvcete biocon!crsion

Based on: A. L. Denain 2000. Microbiai biotechnology. Tibtech 18:26-11: A. L. Demaii 20{X) Phrnir.eltri:it!\ a.l!e scrA- L Dernain. I999. Seconciary metaboiism in bacleria: Antibiotic pathways relnlation and Iunction in Bi.,l,r(r t,! th; pni:at

,nd!^' meebojites 01 microor,ganirms_ .1/r/r. Milrr,rr(rrl..r. J \\'- Len-q.ler G Drcrs_ and Il a;. Schleeel.

I Butahntl. i):155 .{'61: c Lancinredibrs 617-51 Nr\r Yorl: Jhicnr

-l\antlltn i-r()lvntel\ eitl]liltce ()il feco\r.f\, br, trtrl.;, 1\ tnLt \\.ltL,rf'lr)odin-e and the displaceruent ol'oii. 'l'his usc of xaltthun sLul.lll'otlttced b)'Xurttltontotld\ (LuttlL\//t\. tcptc\L-itts ll iarsrt ]t(rtant ul nrarkct 1or litis ntrcr ob jal i)r()duc1.'l'hc cr cl()d.'\irin\ har,e lr uniqLlc \truulLliLt. it\ sho\\ n illfigure :12.1 I 1or cr c],clodextt ip. Ther' are cr clic gliSrr:rictri:r,ritlcs u ht)sc sLr!als iire .ioinr(i br o I .J linkagrs. C-'t clotlcrtiinrcrin bc use(l litl ti uirie \ariet\ ()f Jrul'lloses be.caLi:c thcrc err.ircll ntolccriles hind u'ith suhstances and tilodil'\, tlre ir pitr sicllIlr()pcrties. Iror crantple. cvclode\trilt\ rr jll incrcase tltc :olLrhil-itr ol'phlrrrrurce Ltticals. rlraluce the ir bittentc:is. unal nllisk ( ]tcllrlciil odors. CvclLtdcrtrins als() can ire usetl ir_s sL-lecti\ g 1lr1r1lrg111r

t() r-enro\c cltolcstelol lritrit eggs anri bultcr. t, r []tllcrt \]l IL.\ll orrr oridal ir)n. ()i' us stat ionart' pilliscs in gas chrontitto!i ltltlt,\

Biosu rf acta nts

Biosurlictants are especiallr,. itnportant lbr envir.ot'rr-r-rental allpli-cations where biodegradabiiity is a ma.jor requiremenr. Ther, areused tbr entulsitication, increasing detergencl,. \\,etting and phasedispersion, as well as fbr solubilization. These properties rre c.-pecially irnportant in biorernediation. oil spill dispersion. and .r,hanced oil recoven, (EOR). The ntost widelv used nticrobiallrproduced biosurthctants are gl\ colipids. Thesc cornl(runds h t\ cdistinct hr drophilic and hr.drophobic rcgions. and rirc c\..lldltidispr-rsine agenls.

-l-he-r, have bcen use d \\'ith the l rrr.,r: lrrlajr.-

and other oil spills.

cylpS\-o- /r-v \

s1 PA."q:l^{/) "' i t*Sia*o,bo+ \\'

K,on Ho-,l'o..\ ryo9,o2) )o+_ t/ "t -g o-" 5to) | {[r---,-r- oJ o F

o',AO.HJ

ir-Cyclodexlr r-

Figure 42.11 .{ f'r clodertrin. The \irlrcturc rrl ,,-r'., clodcrlrin.llrtrduct-d ltx I'ltt'prtt,tlrtt(k)j)uLr(,1. Thcrc als6 tue [] 1;,i -v iirrlts. 61'

sinrilar \tniclurc bLit Ialger: these cor.npounds ha\e iltan\ applica-tions in ntedicine ancl industrr,.

Bioconversion Processes

Pr i, rrtr.rir', rl \ ii )il \. :tlrtr k n, r,,., it li. nlicrobixl t ransti rrntations r,r bi<t-transfirrntations..rl-1.;Ilin,,r ih.rn-ac. 111 1111ri3;ulg...r,Jtr ii\ lhr- in-

-\iitron ili .t i:rdiir:.i i trr kr,to liiniti0n ()t tllf \iitLtr.iltl]t)ide\itturaliOn,11'a ci'rrtpler crclrc stnrcture. that are can-ied out br nongro,,ring

L979Microbial Growth in Complex Natural Environments

4217

?'.C:O

Ho- ^. I -l-,i I-l'\J_iMajor ProCuct

Figure 42-12 Biotransformation to \lodifl'a Steroid' H1

ilrox-v-larion of pr,lgeslcrone itl ihc llrr position b1' Rhr:r;7;lr'r rli-

gri,',,,,.r. -fhe str:r,rid i5 il15-'t-'lred in accttlne belor-e ldilititln 1o tlle

ftr.-r,,\\ Ir itlllf :il . ttltLtlr.

n-iicroot-garti:rlr\.'l-llL' ttlicttlt'lgal.iistns iilLls lict li: tlitlcatallsLs'

BiitCorrr e r:tons Itirr e ll-lltllr atlr .intli!e \ i)\ L-l ChellliCill prOCttittres'

A nltjor adr rLrtllige is sterctle ltr:tll jcai' the hirritrticljlr ltclir e tirrrtl

o1' I prodr.ict is rltlttlc. [:llu t'ttlcs lt]str cltrrr otll \ ci\ :i)ccillc rtiic-

tionr utrricr nttlt1 rrrtlritlirll\. jlltl l.it'l't t'.ilut tll'illtthlt tlltllCcttlCs

clrn ht- tftrn:1,1I'n.tcii. l- ni.t1]Lil:il hlrtene. llatill()lt]\aL'tL'\. \L'llsls.

;rntl ntoltl:her..hCtIir:tJin\.1ilr)Li\lr1|q,rl1 -i'l()ll\.'lheCtlzrtltesre spttnsrbJ.. l0i. thc:c !.(r11\ L,f\1()n\ .etl hc liitlltct'llrllllr 0l' t\tflice i

Iuiar. cells 1'1111 hL'1tlrrtli.iteuL jn Lr:ttuil iil !()lltillur,Lis cLtltui.l allrl illcil

dried lirr dire,'t ust-. rrl lllc\ .irrl bc pre pltrc'd ilt tttole sllceif ic uavs

1,, \.,rf\ ',rt1 ,lr'.tr,',1 r'.,'. ,',\s1'j"r''.\ 1)'pialll t iiliritl', 1-l:i(i11 is tht hrdrorr laitilll tr1' a stctorrl

rligure 12.D.l ln this trrtrrpll' thc s arct t"t6lgLrle ste roid is

dissolyctl lp aCL-t(r1c erlii ti'i.'ll ldded tg tl-ie rcactiOtt systell thal

cotttltjns thc llrcurtlrl n rlliclrlbil] cclls' -lhc cotlrse o1'the rrrodi-

iicetiort is tttirrtittlrtri. rlntl tl'tc f inll ;rrotitrcl is e rtraclcil lronl ihe

tllr'Lllllrll llrJ 1',tt tl tr'i.BiOtranqf'onltat ions Cllt rie 11 oLrl br i'lCr' L-ll1)'llie S (lr lntilct ll()n

g11ru tttt uclls tio hltr.' ljnlitatiotls' I{clie liotl' thltl occitr in tlre lrb-

rc,tc" ,,1 uirjrr' ittel:il.olisitl .'r ithtrLtt leclLicin!' llor\e l r)l :\TP

hcing ar arlablr- a()lltllltl3ll\ - ilrt pt itnal'il\r cxcl q()lllt tcitL tir lll\

(sr'(' .t('( 1lr)i r ,t -j i. l1 {l-P ol rdrlLlct:lllts rile letltlired' sn cnerg\

':ource \Llch 3 gl111 ose ]lltl\l ilt'sirpplied urlcleI carclirlll'eiln

tr olletl ttotttr()u th arrlltlili()ils

'l Discuss the maior uses for biopolymers and biosurJactants'

2 What are cyclodextrins anC why are they important additives?

3 What are bioconversions or biotransformations? Describe the

changes in molecules that result from these processes

MICROB:AL GROWrHIN COMPLEXNATURAL ENVIRONMENTS

\,licrr,hial ntzlnaltrntcnt ul\(l ci]lt be carried ()ut in colllple x natu-

tll cnr !;o111llr'llt: -Llail ii\ \\llt!-r(- st'ils' 0t high ''t':'rnic lll'rltt-r

conlillrlin! aLlillPrr\i\ u lltrt- tht phr:tcll 'rtttl ntllrliiilllui

conditiirn: i,r r1ticr,,bitrl -{i()\\lh (11n1()I be col1iPlLrteiv contrtilled.

aniJ a largclr Ltnknolrn tllt.ittbt.tl rrrll'lnlunit) i\ present. Eranl-

製ilcul」ねFli:だ11lliF[Ft札獣W‖llnl猟轟Ia鍵

附f凛 1薯 11風)lin」責1撃。∴篤H∬蹴∬t選尊plicationsヽ 11l bc discusscd in this sCCtiOn

31ode9radation Using Natural MiCrobialCorllmunities

IPIittF‖1棚新ざTTi朧瀞ぶ緊i諄寝黒:

prよ iou、 l、 (｀('('/'59イ ).nlincralization i、 lhc llanSfOHllation o「

()rgを llliC nlolcculc、 lo 11111lC'cti l()1■ ■ヽC()litillall、

itヽ tlS assu11lcd gi、 cn tinlc and thC a11110st inl｀initc

ェI:や|1鍵:1罵gギ寵:キ1llti11:|=ポ ilまま:1属li鷺:lT

Oい、cい ltiい n、 。f na■ 出 i mdヽ )nthctic orgtti… ■甲 ■nd accu―

1lRliatioll i1l ntlttll.llビ l)ヽ'il()nnlclltヽ .hOヽ cヽヽ cr 卜c=an lo ralヽ CЧヒIC｀ ~

ti()n、 ュ1,o tlt[l〕 c tibilit卜 ol｀ nlicroo「=ani｀

111｀ 10 dCgratlc(hescヽ tll ic(1

、ub、 ltHlccS tlll(l thC lolc()i thC c11ヽ il ollnlclnt{CItl卜 、 allaCrobic c● :l「

‖1墓1驚[‖ili鮮湛‖i鮒黒1灘‖淋l醐i鸞肺=思

(‖

織:I里‖謝ド(濯#寵l:起II‖ ll。 lliふ「llよ1よ

週lIIli椰‖IIt卜iぎ1織IIIII11螢 1戦[liヽ c(11「o lllヽ aヽtcrl othCl tヽtldic、 llndicatc th.lt h)drofcn cti11卜 c thc

OヽtllCe Of lctluctを ll]t i10r thc dChalolcllation of(liffclcl■ C111011-

1111ダ llII常 i′)|‖;1淵 11ッ‡||′ }‖lぶ オ ヽ`器

下|)力l

Hunnic iCid、 .br。、Inish polynncric lCSiducs t)f lignill dcconl

驀 輔 幕 鮒

md:鰹 °

鷲

JttlHc醍崚 mmm囃 ぃ rC mmJd測

dc gi atlati(), ol thc lllを li 11 、(1■ lCtlll C 01'nl(ln)pc、 tiCidC、 tlin、 l ollnC:

i鼻彙!1熙榊‖illttIIiI瀧iII珪

Chapter 42 lndustrial Microbiology and Biotechnology 4218

' 42.13 Biodegradation Has Several

ngs. Biodegradatit)lt is a iernl that can

I to cle scribe thre e rna.jor t1 pe s of changcs

rlecule. (a) A minor change in the func-

lroLrps altached to an organtc cot-t'tporrnd.

substitution of a h1'tiroxyI gloup filr a

e rroup. (b).Art :r, iulll brr:aking trf ihc or'

onriround inio orgattic fiagtllellts in sircil

hrrr rhe original rnoiecule could bc reciln-

.i. {c') l-he conrpletc clcgradation of all or

ompor.rnti to rninet lls-

(a) Minor change (dehalogenat;on)

Che ln ca structt,re

O~~l~||~C00H

24-D

G~~CH ~C00H|

α6……… …H6…(b) Fragmentaiicn

G6……… α6…嘲

(c) Mineralization

.ciGO…剛―…翻

Approxinrate time todegrade in soil

3 months

2-3 years

81ocked″ ,era pos tiOn

(b)

42 14 The l′ εrα Ellc・ ct and Biodegrtldation. 1ヽ11lo「

ll di■ cl cllCCS CtlI〕 hを 1、で illl10r CllCCl、 ぃ11 lhC bio(lcgl」 (1-

)1'chenlical、 Thc ″′てィ`′

Cticct iヽ ti:〕 inlPo「 ttl1lt Cヽ そl11lplc

」il)i dcgradablc 2、 1-dic‐ hlor()phcno、 )'acctic acid (2、 4-D,

cxposcd nlcta position oll thc ring dcgrtldc、 111 、cヽ 'Cr[J

(b)rCCalcitrallt 2.4.5-trichlorophcll()入 卜'aCCtic acid(24.5-

thc bloc‐ ked nlcta group can pcr、 1、 [forゝ c aI、

。PPO｀ Cd t。 11lc● 1lh・「

い、iti● n ti〕 ビc(ヽ :l]Pい tilld｀ 1`1l l‐ ごcic―

l ti nluch、 1()、、c「 rtllc ]hc″′`ra effect i、

、hlヽ 、、〔l i1l Agure

hi、 、tcl cぃ chClllical(11「 lcrcllcc i、 lhc rca、 on thatthc cぃ 11]―

nrl)rt lir\\ rt it,-'ilri.i.lr i.1 tll.llJrrl',r1lh.'ntltr ile i1l. iif ill :l.j-l)t.uitlt lt cltlrrr,.r. ir: thi,r11111r Ir-rrttirrn. trll hc llrrg-elr Llcrllrie.l inlL singlr \unrrar lil Llrrlrli:- 1. i.5 1t'ichltlrrlDhctlo\\ ileetic ai'itl.\\rthilri)n\tiill(rrllrrliiriIllr-tlllli)\i1i! l.rri11lltlrjstirltlt.'.lilslirr'\r,\ L-lirl \ cill \. llnrl litu> t: i-r:cd irtt ltrt,:-t.:ltt lrt rtsit e itttttol.

.\l inrpirr.illtlrl:Ir.l ititttlrttltgrttL lrlrl,-lcSterilillr.lrl ts tltc t-ecr.lg-

nitiort thlit IIltn', 01 ihi'etltlllloutlds that atc arltletl to t'llrirott-nreirl\ llrd chiral. (rl Ir()\\ess lls\ nttllL-1r\ lind Illttltledtless.

\ii.r',roILlittirltt\ ()ititt ilttl dcg|ltr-lc (ltll\ (llle i:otttct "i e ttti.l-\tltncC. tire iltll-r i\()ntcr' '," til tctlluirl ltl tlle ttlr itttttllletli. ,{i lca.t

l--it, r,l ht-tl,iii,1t: ltre.hillii. ThLr. rt i: ilillcal lrr ldtl li-re nel'bi-

e itlc irortrcl lhiit l\ c11..ti'. c ltttd lLi.i, de grltdliirl.'. [(uct':li t'.ttdtr's

irer c -ho',i n thut nricrohtal ctrltttitr-tt.titie s irl dillert'nt e nr iLon-

nre nt\ \\ ill cirtgirrcle rjii'te lcnt ellrttltiottlcrs. Chltll-re . itr .ttt irotr-

nrrnt:ll conciililnr airtl nutricltt:iipltiics c:itt rtltet tllc petterns o1

i irilltl l olnt rl.-.! tildlili(rlt\1i.r,;bi.rl .i)nlrllirtilics cl.tltltg.- ihr'ir chltlt teri\tl!\ lll IC

\ll()n\t ro tllt- lrrirlitirrrl,tf iilrlrgellie ',r trrglttlic -iLtllsll-lttcs. [1'l par-

liculel cLrnrptrirrrl. \uch li' l herhicttlc. is.rkler.l rr'Peaicdl\ io a

nriclohirrl crrrrnruttiil. lhe citt-tttttttrtit\ 3dili)ti and fastcr rates l)lilegre.letion i.rrn ()ccur a pr(lcLr\s oi licclinleiiorl (figure 42.15t.

The aiiaptir L' Irr().e\:i o11tn is so ci'1'ectir c tilrrt this cnrichrtlent

culturc-bes.'ti ripprouch. estahlished on tite Principlcs elucidlted

b1 Bcrje rinck \.st,t' p. l/ ) can be Lised to isolate organisnts r"ith a

dcsir.-d se't ol.crpabililies. For erantple. a microbial comntunity

can heconrc so cfficient at rapid herbicide deqraclation that herbi-

cidc cl1-ectii e rtess is dintinislted. To coullteract this 1-rri-'cess. her-

bicitlc. clin he changed i(r tirro\\ thc nricrobial ctttittttunitl.ollblllrn.:. ihrl. i'r i':ar\ ilt! ihc .'ttcitir t'llr'Si o1' tilt cht-rlliceis.

i)t'gi:il:tlt,,rt iri{iar'\.a\ ihat,rcCttt in rtlll. llso clitt L-.c uscd illl:rtSr' :-lric J:,ii.r,l.riiLrtl ir1'hrrirtlcltti,)ll\ ()! \I'3:ta\ ti(illl iigl'tcui-

tLllal opL-ratl(rris. in a tcrltirquc talleii land farnring. The uasle

Microbial Growth in Complex Natural Environrnents 9創4219

，Ｃ●ｏＬｏＯ）

ゆＣ一Ｃ一ＣＥ

Ｏ」０つ一０一０」０エ

Time

Figure 42.'l 5 Repeated Exposure and f)egradation Rate'

.Ariilition ot att herbii:ide to rl soil cln resull in changes ill the

dcgradatir e abiiitl' o1' the nlicrobial cotlltl.ittnitr'' Relative tlegra

ri:rti0n ra{cs lor an herbicide a1'1er initial lddition to a silil. antl af

l('t I('lrc:llc,I c\p{)\tllt lrr IItt rittttc .h'ctii'"rl'

Figure 42.16 lヽicrobial‐ l■ lediated 卜ICtJ Corroslon. Thc

lllicr`ヽ biol()tical con osi()n ofir()niヽ an]lo「 p10blCin Thc=raphiti―

zati()11()fi「 OI〕 undcl a nl、 l blcb()n tl]c IDipC Surll、 cc tl1lo、 1ヽ1liCr001-

gani、 ■、 i1lcluding Dど 、′``rr l〃 '′

ル,toこ ()ロス)dc tlnc inncr sulfacc

The nrethanogens. an inlpo(ant group of the archaea that can produce

metliane . har.e had ri'idcspread impact on the Earth. Dr'\pitc intensir e

research. neq' discoveries are still heing made concerning lhese nli-

croorganisms. Methanogens have nou' been found to cont'ihute to thc

anaerohic corrosir-rn ol st.rft iron. Previousll' the microbial group usu-

ally consiriered the rriajor ctllplit in the anaerobic corr)sion ploce\\

r,,as the renus Dr:-safbi iDrio, lvhich can use sulfate as an oxidant and

h1'cirogen produced in the corroslon process as a reductitnt'

Nlethanrtgerrs cail ilse elemental iron as an electron source in their

metabolism. It appear-s that corro'\ion mev occur even rvithout the

pre -qeircc o1 sulfate. ri hicil is requirerl for functioning of Desullitvib-

no. Rates of iron removal b1 the methanogens are around 79

mg/1.000 cm: ol sutlace area ln a 2':l-ht-'ur period' This rn',u nt't tttn'a.lugh rate. but in ir'lation tl ihe plannetl service life of metal struc-

Iures in muds atrLJ suhl,surl-ace :olls-po:sib11' l'ears and decades-

such corrosion can become a major problem- Continuous ellt)r1s to

improYe protectlon of trtrn structure-t $'ill he require'J in vie $' of the

r-lirer.itr ul tr.'n ., rr, dinr iili( rooT-:.ir:'lll'.

nrate rial is incor.lturated into the soil or allo\.'efl 16 ilirr,r' ltct-riss [he

soi1 sLrrface. u herc rit-gratllttiotl occurs.

It is intportant ttl enrphasizc il)at such degradation Pr(lcLrsses

tlo not alrr a),s reduce en!ironntentnl problerrrs. In I'act. tlle partial

degradation or modif ication of an organic compound rna)' not

lead to decreased toricitl. An exanlple of this process is the rni-

crobial nretabolism ol 1.1, I -trichloro-2.2-bls-(p-chlorophcnl'l)

ethane (DDT). a renobiotic or foreign (chemically s)'nthesiT.ed)

organic cort.tpound. Degradation relnoves a chlorine tunction to

gir e 1. I dichloro-2,2-bls(p-chlorophenyl)ethylene (DDE)' rvhich

is still of environntclttal concern. Another irnporlant exanrple is

the degradation of trichloroethl'lene (TCE). a u'idel1' used solvent'

tf thii is dc-gredcd urtdcr atllerohic conditions. thc dange rotts car

cinogen vinr I chloride can be * nthesized.

■Diodc,Ind(ltioll al、 、、 ご(11l lCti(1 1い 、、itic、 1)rCをld dalllagc、 a11(1 11-

nanciallぃ 、ヽcヽ lヽctttl c■ 11(｀ 1ヽい ll i、 tl l、 11liCtllal l)i1111)。 11を11lt Cヽ anlPIC

Thc l)licI()biallヽ nlcdi(ltctiぜ い1■ o、 it)I〕 ぃ1 1llCtttls i、 I)a]■lcula11卜 ごril―

ictll、 、hcrc iroll pipc、 1lrc tl、 cd il]ヽ tヽitcl l(,g=cd t11lacrobic C11、 iron―

nlc13t、 o: in sccondar)pctF(,lcu131 rccい cヽrヽ ProccSSCS CarriCci c)ut at

oldcr oil llcld、 IIn thcsc r)ldcr llcl(1ヽ 、、をitCr iヽ pu11lpcd doヽ nヽa、 c―

lics of、 こヽll、 to l]orce rc、 idヒ lを11 lDctrい lCtinl t()a CCntral c()llcction

point lf tllc、 'ヽatcr co13taiI]、 lo、 lcヽ cls of organic(1lattcr and sul―

fatc.anacrobic 11licrobial co:1lnnunitics can dc、.clop in l■ lst blcb、 ol

tubcrclcs(figure 42.16,.rc、 ulting in puncturcd iron pipe and loss

of critical ptinlPing PrCSSヒlrc lヽicroぃ rganis111、 that usc clclllcntal

iron as an clcctron donor tiu〔 i11=thc rcdtiction of COc in■ letinano―

gcne、 i、 ha、 c rcccintiヽ いccll di、さいヽ ci ctt (｀ Iicrobial Di、 ersit》

Ecolog}42.2)Bぜ じ(ltl、 せ●fthC、 lヽdき「こ:1=C｀ )「 intcractit)ns that oc―

ctlr bct、 ccn 11licIて 、oritini｀ l11、 tl■ ご l、 iciユ lS thc nccd 〔 dcヽ cl(,Iつ

$tratcgics to dcalヽ iヽth 11licr(ぅ いial c(ヽ 1l oヽ 101l PrOblClll、 1,clticalClr - CHC1 -+ CIIIiC - CH:

一 initial degralation pattern

MO●org甲 1甲 ,W“

'9ut

“CめⅢ籠Sms＼ :船董1蘇"側

Юb

出鍛ぼ＼■||,‐・‐■|'■ |||:

「

h4 = Monitoring wells

:|||

Figure 42.17 A Subsurface I'-ngineerecl Biurenrediution Srstem.iiirtl it: Jro\sil)lr nl()\cnrcnt. \utrie ntr and (l\\ ge r) (ii\ iiir. pero\1dd. ()r

!r'(rLlnci\\ !ter 1(r ])rOnlotr- nrLtre e liiciclrt i-lcgr':rdati0n r.rl'crtntlintinltnts.

涌ヽ /1｀/1｀

/1ヽ

=Contaminants

Itiirrritolin:, lrnd recoverr u,ells rire u\cd t(l nrtrnit,rr the plumeo1h.r ()\\ qL-n rclL'a,qing cornpounds) ar-c added to the soil and

Give alternative deiinitions for the term biodegradation.What is reductiv€ dehalogenation? Descri-re humic acids and

the role they can play in anaerobic degradation processes.

Why is the "mela effect" imporlant for understanding

b iodeg radati on ?

Discuss chirality and its irnportance for understanding degra

dation effects in the environment.What are some of the irrrporlant mrcrobial groups rnvolved in

iron corosion?

Changing Environmental Conditionsto Stimulate Biodegradation

Olten natural nircrohial conrnr,-uiitres *'ill not be abie to calrv outbiodegradaticn proces\e s at a desircrd nitr' due to iilniting pirlsicalor nutritional factors. For e rample. bioctegradation cilien uill be

linrited hr, loq ori,\.!.ll lercl.. Hrdrocarbons. nitrogc-r.r. phospho

rLtq. and othe r ne.'rie tl nLrinari\ :il.r, rnlrr he absent 0r ai arlablco1llr 111 rl11u. flLrr rti.r.. iirLi' rr:rrrtrnt iJia' {rl dr'rr:idllilt)it. In thc.e

alr\c\. rL ls nL'ac\siir v kr tlelclrnine the Iinriting 1ric1()r\. hlred or.t

l-irbiu''r lrnrl S jrelford's la* s. and tircn to sup|lr needrti nr:rtelials

or.nrririilr thc cnr ironnrcnt. i . . .. ,, ',i:, r ,i : . ,

\lr)-\t ()1 tlrc clirlr ctlorts to stinruiate thc dclra(irti,.e iictr\ i-tie: ol' rnie roor-guni:n.ts irrr olveil thc nrodiircation olr \\ aters and

soils br thc uilditron o1'()\\,gen ol nutrienls. calied bioremedia-tion rrnd slroun iri figure -{2.17. Coutact bctuccn tire nricrobcsantl thc subsllate: the pnrpel physical envilonnrcnt- nutr-icnts.

o\\'gen { in nrost clises): and the absencc o1'toxic conrpounds are

critical rn ihis rrranaged process.

OIlen it rs firund that the rddition of easilr melaholized or-genrc rnatter such as qlucose incrcases biodegradatrr-rn ol recalci-lr'ant compounils that are usuallv not used as carbon and ener-et'

sources bv nricroorganisms. This process. temed cometabolism,is frnding rvidespread applications in biodegradation manase-ment. Cornetabolisnr can be carried out by simply adding easilycatabolized orsanic matter such as glucose or cellulose and the

compourrcl to he degradcd to a corlplex nricrobral comrriunitr'.Plants also nral he used to pror,ide the organic nratter. C{lnrdtaholi:rr is irnfor-rlilt in rran\ dilterent birrde-eradation s\ \ie nt\-

‐■理

4

「

4221 Microbial Growth in Complex Natura! Environments 983

弊 C02

Process F unctiorl

Stimulating Hydrocarbon Degradationin Waters and SoilsEr;rericnccs u,ith rtil 'ttjll. rit itlllirla clli ilrttllttt'llls illlt'ttllttthcst principles. \\iltert uotkitlL rrllh tli'llcr:ctl hr,ltLrcrtlhoirt itl

the occan. contllct bet\\ cctt tht ttticrooiLltnisrll. th.' ltr clr,-tcltrl-ton

substrate . antl othcl e sse ntiJl ttLtllicltts illust bt' ttlltitllltitle .l. Ttr

achieve this. |3llg1s t()rlttlitrtll-! lrtllri.nt\ llrld iill 1r1.1)Jrhilic ihrdrocat-bttn stilLrhle') l)icpilrirti()rt lllt',t hecll Lrs.'ti -1-iri. lrthlilri'.1.Itas acceleretcci the tltslitlrtiltt,rl rliiltt-titt ciLti1.. r,tl .iie k. br .:()to'109i . in crttttpat isrrtt ti ith c()ltlt'rrl t,il :ll.k. u ilelr lllt lttltliltottltlnutrient,s \\'L're not lrvai lanic.

A nlljor challcnge ior thr' 1slll1rlirr1r r'..tr 11lr' l:t'"tt \rti,lr''ciil sprll. u,hich occurrctt ilt .'\llr:k:,r irl \llirch l()8(l Scr crltl drt'

tarcnl allploaclles \\er' Li\i'(l t() itt'tt'list bir"lr:!t;r'lrltion Illcrr'inclLtclecl nutt'je nt additions. chtrrticll d j\lt!-!'\ltttt\. hi()\Lll iil.tlltllaclditions. arrri the use of hicit Jrrr'\\tttL- \1..trll T]1c Ll\L'(i1 il rlli-crobiall)' prodrtccd Slr colif id crlrul:i1'i.'i i.trt. irrt'r trr helpiLll

Thesc bictretttetlieltott;t|1ll-i-rltihr'.:il'a ll\!r LlscLl itl 'Lrils lLllti

seditlents. Filreranlpl.'.:i Illlirltlc r\\r'|-\t.l:lc J\l ,rrtr\\ tltn ltt Lt'ed

to degrade PC'Bs in riret .cdittte nt:. Irirst. piiltiltl tlt-llltlogcnilti(rllof the PCIBs occur\ nalurallr urttler ltnrL-r(lt.riL trrnililioils Tilen

the rnuds are aerated 1(l proittolc tl.tt- cotlrplelc de gllttlaiion tlf thc

less chlorinated rcsidtie s ie haptc-r opening figurc r

Stimulating Degradation with PlantsPhytoremediation. or the use ol pl3115 to stinrulate the ertractitln.degradation. adsorptitrtt. stahilization or r.olatilizrttirrn lrl aL)lltllrll

inants is beconlinS an inlponant pill't (r1'hii'tleSr:lJ,rtj,'n rt','htroi-

og1 . .A plant proVidcs nulriL'llt\ th lt '1ll()\\ rrrlllt-i:ih" rlr'|l r\ "t.Lrrin the plant r(rot zonL' or ritizrrrpilere rfigure 11. I E ,. Pllr i,rlcrllcdiatinn lll-\g inClude:1..liint a!tltrilrLtIj\rr.t\ l(),ia"t,1.1.1::r,ri. i:-llitl,rhi

■■

■■||=

こ え

CI CIGOGCI CI

ing t. enhanCed dr:gt-adlrti0n rtf cOntnrrrrntrnt:. Thr tlrgftttllrtion rri'hcrltihlLlt tlbetlzctlt is shilull !: llll Cr:tlllp1t.

Phytoextraclion

Phytodegradation

Rhizofiltration

Phytostabiiization

Phytovolati Iizarion

Llsc 0l pollurlnt accuntul;tting planls to removenrelals or orgrnics iioir soil by concentratitrgthenr in the harvcstebl!r plant pans

I -- | 1'l ril'' :r: 'l r.. .. ii''CJ l' lc'o 'l:alll\m\ Ir'

dcLirrili,rrgliric prll uiants

Lrsc of pllrr i|r)t\ 1(r 3Lr\(lrb uld ad,.orh poilulilnts.

irrrinlr r,rrtnis- 1-r(ri| \i'iiler and equeous rvaste

stIeaJlt \L.\. 01 lllen1s irr reducc lhe bioavarlabilitl'ofpollulants

ln iire e:rr LaorilcrL

Usc ol planr: io r',riatilizc follutar.tts

Based on T. Macek: N'{. Mackova: and J Ktis.

in cnlironmental rcm&lialion. Br.rl.drkrl. .4/lOЮ ExPloitation of plants fc・ r the κmoval of ottanics

18:23-3■ P 25

lizをltion ancI 、:()ltttili/tlti()11 lDl()CCヽ Cヽヽ まヽ :n(ltCd in table 42.12.

Tl a11、 gc:lic PlをlI]ts I1ltl卜 |)C Cl]]ID()卜 Cd i1l Phゝ lorcnnediatioll Usillg

cloning tcchlliЧ tlc、 、ヽlth マ`′ャ,わこれ′ど′ll`′ ll(ヽ どで'ノ'ノ '「

ノゞ, イア9-Sθ、

/10(').thc″7(′ ′へを11ld′ 71て '′ B gcncs htl、ビトccn il〕 tegratcd into a plant

(1′`7/,た

ノ`,ノ

,、 ′` 7/7`ノ f`″ ,て

`)thtIS illをlking it po、 siblc to transforill cx―

tl‐cinCl)ito、 ic()rgallic lllcrcur卜 f()「 11lS tO Clcnncntai nnercucy,v/hich

is icss of an cn、 .ironnlcntal ha′ ard Rcccntly transgcnic tobacco

pla1lts ha、.c bccll cons(1‐uctcd that c、 Prcss tCtlanitrate reductasc,an

CnZ卜 ]1)e fl olll(1ll Cヽ Plい 1ヽ｀ ビーCicfra(Jing btictcrit111l thcrcby cnabling

thc tran、 ビclliti l‐ ]ttll t、 1、 dヽC」「t,tic nilltltc e、 1ご r and nitro aroinatic cx―

PI()｀ 1｀ C｀ ThcゞごInご t:C ti:l卜 i、1ぃ (ililCd IPItull、 rゞO｀ `in s()lutions of cヽ ―

Plい 1`｀

=｀thtttご ぃntI Ⅲl:ヽ itinl｀ Ctilinて tヽ llヽ 1ぜ Itiビ (Dthcr Plants ha、 c bccn

Phytorcmed iation

Chapter 42 lndustrial Microbiology and Biotechnology

Figure 42.'19 Copper I-eaching fronl Lou'-Grade

Ores. Tlte chenri-strr rrnd rnicrobiillosv oi'coppe r ort'

leaching involre ititcrestittl clrllPlelnentlil re:lctjons''l-he nricrobial contributioir is the oridatitln of lenous

iirn ( Fer' ) ttr flrric irln (Fer - ). LtptrtsTtirillrrnt.fe rrrt'tt'i -

,1rltr lutti relaled Inicrrltlrg.rtli:ttlt al't \L'r\ lctire in this

oridatirtn. The terric ion then rcacts cilelllicalll' to sol-

Lri-.ilize thc copper. The solublt coppr:r is reco"ercil t"

l chenrrcal reactiorl uith elcnlenlal inrn' lrhich rcsulis

r n :rn ('lrrIlclitirl .,,|1rq 1' fl rt ;Ii1111,'

r'nginr'elcrl in the s;Ltllc \\l\ to (lt-griille lrichltlr'rt-tht icttc rtl ''lt' i

i-r)n IIlL- rll ;1 I aol-lIll ll I i llllilt i li $ tlrl dlr t tle ctlt.ict'l tl'

Stimulation of fu'letal Bioleaching from MineralsIlittleachin\, ts tlt.. usc of ntic|o()r'gat'risms. u hii.:h pioilLicc rtcids

ironr reducecl sull'ut ctlltlllot-ttl'-ls. lil crcate acidit ctlr itulttlletlt'

that sttlubilizc desir.eil nretal-s lor i-eco\ cr-\" This apprrlaclr is tlst'tl

to reco\,er lnetals front or-cs ltncl nrininq tailings uith in.'tal lcrel:

loo 1ou, 10r ,cinr,lting. Bi0ir:rching carritcl otri bY natulal popLrla-

tiotrs rrf Lt'lttr,.sTtLrillturi-iike specic-s. Thiobtr'rllttr 1/lirrrr'izrlrtrtr'

encl rrlui..tl thiohaciili. l'orerattlple. ullo$,s recoierr of up to 70'i

r)i thc cr)ppci itl lotr gradt' orts. As siltlu n irl figure '12'19' this

urr olr es rhe biololicel ,11id'11ott ot'ctlppe r prL'sellt in lhe se ort-s

to producc solLrble copllcr srrlfate.

It rs apparent tltat italllre $iil assist in blorc-rllediation i1'riVen

a chance. The lolc tlf Illtrul-lli tttict-oorglttlisrlls lll biodcgradation

is nou hetter-apprccirietl. Arl excellent eralrple is thc receni

,,rtrrk.,,,ith the ren ret-slitile 1'ungus Pltattarrtt-Jtat'tt'tltnstt.slttt'

i.rrrir (\[icrotlial l)iversitr' & Ilcologl''{2'3 t

Ol.tenhiodcgr,.lllilrnlndlriodeterirlrationhalt-Ill.ti{)rlle.Jjtire citects. utid it becoiltes intportant til ctlntrol and lirrlit thcse

processr-s b1 e n!irotitlle ntal mallageurent Problenls include ull-

,n ant.,l degraclation of paper. 1et 1'uels. tertiles. and leather goods.

A global colrcern is microbial-based metal corrosiorl'

1. What is cometabolism and why is this important for degrada-

tion processes?

2. What factors must one consider when attempting to stimulate

the microbial degradation of a massive oil spill in a marine en-

vironment?

3. Describe the major types of phytoremediation What is the role

of microorganisms in each of these processes?

4222

Precipitationo{ copper

FeS04

4. How is bioleaching (arried 0ut and \'vhat mi(!-sbiai genera are

invo lved ?

5 What is unique about Fi:aerachaeie chrt'scspltrLtfil? What

does its name mean?

Addition of ttqicroorganisms to COmplexAllicrobial⊂ onlmunities

B(ヽ th i11 1(1い ぃrtit(ヽ 1ヽ alltl liel(1、 t ti(lic、 tittClllI)モ 1`lttヽ こ卜ecil llla(lc lo

、pcc(l t11)c、 i、 tinピ 11licIい bit)lい iゞCtil l)Ioccヽ Cヽヽ 卜〕こ_ldil,ピ kn()｀ 1`1(lC―

ti、 c llnicl otlrtalliヽ 111、 1(ヽ (ヽう ilヽ ｀`[ltclヽ ●l いllncl C()1111〕 :き `、卜ヽ tc:l'｀ [1ヽ

|ボ尋置I1111‖鷲‖》][||:1驀li三:椰1llcnt culttllcs For c、 tinlPIC COnlnlcrcial ctilttilc I)「 CPttratlo11、 arc

tl、 alltlblc to lhcilitatc、 llagc lolnlを 1lion tllnd io i1111,1い C`｀ Cptic tank

PCri'()「 11lallCC

Addition of Mi(roorganisms、″ithout Considering

Protective A/1icrohabitats

Vヽitll thc deヽ cloP1llclnt()1(hC Supcrtう tlゞ

~bゝ=｀ 1`(11lakrabait)in

1 971 tincrcヽ aヽS illitial c、 citc11lcnt dtic lo thc hopc that such tlil illl―

proヽ cd llnicroorganisnl lllight bc ttblc to dcgradc h)'drocarbon

囃 lTR:『 1∫ I:|:書 i::詰∬ 1111tl∫ lattLih∴彗∫‖cC‖

crobc had to sur、「iヽ c and functio1l Enginccrcd 11licroorganisl■ s

、、lcrc addcd toヽ oils andヽ atcrsヽ iヽth thc cヽ Pcctali(♪ n that ratcs Of

dcgradationヽ o■ ld bc、 linlし1latcd aヽ IhC、 cI]]iCIい Orsa1liSnls cstctb―

1lshcd tllcl11、 cI｀ Cヽ (]cncrtlll)such adllitit)nヽ lcd l■ hヽon-lcrnl ln―

crcascs in rate、 ()f thC tlcsircd `ご tiヽ 11) 卜ul i卜1ヽicilil artcr a tc、 、

da、's thc tllicr()bitll ct)11lnltinit)Ic、「t｀ 13｀ Cヽ ｀`きrtヽ ::ミ ,il〔 ir ill trcュ tcd

CuSoo + Feo--> Cuo+ FeSo.

2Fe2｀ S°J3+CuFeS2+2H20+302-'CuSO`+5FeS04+2H2SO`

＼

The basidiomycete Phunenrclutete Ll!.n'sosPori tr? (the scientitlc name

nreans "visible hair. golden spore") is a fungus rvith unusual de-grada

tive capabilities. TIrts organi-rnr is terltred a "i|hile ro1 fullgtrs" because

of it.s ability to degrade li-snin. a randonl-v iinked pltenvlprop'ane baseci

polynrerrc component of u ood (se c .sectittii 28 -l). Thc cellulosic por-

tion ofwood is attacked lo a lesserertent. iesulting iil the charactens

tic white color of the degraded wood. This orsanistn also degrade-s rtruiy amazing range of xenobiotic ctmpound. ,nonbiological foreign

chemicals) using both intraceilular and ertraceliular enzvnles.

As erampies. the iungu-s degradcs hetlzt-nc. toluene. ethviben

zene. and ry'lcnes (the so ciilleil B-l'EX conlporriltls). chlorinated

compounds such as 2.-1.5 trichlorocthl'lene (TCEt. rind trichlorophe-

nols. The latter are present as conl:tlllinants in lr orrd prescl i :ttil e: .intl

also are used as pes{icides. In addjtion. 'ltir':: chlrrl illllied henztilts

can be degraded u'ith oruilhilul 1()iLlene! beitlr plc-r.'nl F't'n tllt ill-secticide H-rrdratnethllnon is degratledl

Ho.'r does thrs ttticrLrtrrganisnt carrr ()tti \uch ttllt:i \1.plre:rtlr

rnosl dcqraaleliott oi'thcse xcnilhiotic alrllln(rurld( (\L.Lir' ,,! .r .ttll'' c

growth. during the seconciarl metaboiic lgnin degradation phase.

l)egradation of some contpounds inr olves important extracellular en-

zymc.s inciutling iignin peroxiti:ise. rnanganese-dependent peroxi-,lase. and glyoxai oxidase. A critical enzlme is pyranose oxidase.

ri hich releases H,O. lor r-rse br, '.hc ntan,uanese-deprndent peroxidase

enz\ rre. The H.O2 also is a piecursor of the highly reactive hydroxyl

radicel. which parlicipates in u'ood de-rradation.,A'ppa,ently the pyra-

nose oxitlase .nzyme is located in the interperiplasmic space of the

tlngal cell rvall. u.'heie it can functrott either as a part of the fungus

or be reicased and penetrate into the ri'ood -rubstrate- It appears thal

th!' nonspecrflc enzll.natic s,vstenl that reiease-' these oxidizing prod-

ucts degrades rnany' cyclic. tromatic. and chlt-rrinated compounds re-

lated to lrgnins.\\'e can e\pect to continue hearing ol manr nerl' acivances in

i.ork gith this organtsnt. Potentjali! valuabie applications being

studicd include grou,th in bioreactors rvheic' intracellular and extra-

icllullr enzlnres can be iraintained in the bioieactor lvhile liquid,.\ i1:r.s fl(\\\ pirst the irrmrtbilized fungi.

litltl cirlllrill \\ siallls. .\1lt'i ltlllllr tlll:il''l't'\'1Lil :1111'lllIL\ li \r' 'i\|,Lrnd thur thc ieck o1 cllectiretla:s ol 'L'lll ltldad ctllltllas riar

rlttr- 1O et least thtee lliat(rr\: ( I I tila llltlrtctirelre:' o1 l.rLrItltlirtr

!l0u n ntictrli)tlillllslll\ a: a foiltl \(rtlrae 1i)l |ladlltol': \Llill it: \\)il

Irrotoz(lir. ilt thc irlehilit\ ()1'tht-\t ltticic-d tllictilrlr!.inl\llt\ llr trill-tiicl tht .onrpouttLi\ to he dt-glitil.'d. lrlil il I the iailLrr.- rrl thc

ltrlilcd n-iicr(tol!attisrtts Ii) \Llr\ ]\ L- :illLl c(rlllllctt' q ith intli-gt-rlilu:

ntieroorglini:nrs. Such a rni-rrlii'icLi tllictoirtg;illlstll tlllir tr.'lts' f illo a()nrllr:1a lutd srirr i"t hccartsc trt'tht lttiilitionltl ilt.'ltt-tti hltr

,1rrr rcqLrirttl t() llllutttltilt thc crillr D\.'\.\tterlllltr hai e bcctl tllaclc ltr tllltkt .it.h l.lhr)l illt)1 \ !rr)\\'ll

.ultuie: rll()ra ailf:Lh]c o1:ulrir.ll ill ll lllttlllill cll\l1r)nlll!-llt h\

Snrrrjpg thcnr itt l(r\\ llutriellt tllttlili t.r sIltliiltL tht ]llitJilrrtullnisnr: be lorr' arldint thcltt ttr iirt L'tt\ il()lltlleilt 'l'ila\e tougllr'ilitlt"Irppr-rilcitts hltr r- itttl-.rpr tci Itlicrobiltl sltrr tr ltl ltlti lllllcli6tt slnle-that. bLtt hr1\c rlol s,,1.'.'ii lilc ilr(lhlL'lll. Ill Ir-ct-lli \clil\. tildrd hli\

i-rt'lr les: intcre\t rrt .itllplr lltldill-! llllrrool!lltti.ltts I(' r'll\ iltrll-nrcitts ,,\ ithout cort:rcitring tht rpt.'illc tlichr ol 1111q11rq'rlr it"llnrL'ntinuhichtlt.-r ltl etostttritc:tiltilirllctiotl. l-llislllislcdtothtlitltj ol natural attenuation. lr hich elllphltsizcs tllt- tl\L- l)l lllttLl-

r ;rl nriclt,bial conrtttutritie. in the tttvtrolltttctltll ll l'lll-I!clllcllt'i r1

poJ I utants.

Addition of Microorganisms ConsideringProtective M icrohabitats\'licroor-canism additions to ttatttral etlr,irotlrrents ciit) be nlore

:uccessful if the nticrorlt sallisnl is arlded together rl itI-t lt illicrtr

hribirat th.t lire> the i)rgani\lll phrsicll P1{rt'cti()ll':is *ell e:

irr:::ihlr \Lrlpl) il'jg ntllllL'lli\. Tili. ill-ik.'. ;i 1lr'::li-'ie ir'r lilr- illi-

-:,,,,1-tLrlr:nr i() \r1r'\i\e in s;ltlc o1 ill.'itltcn.t'a()lllpctitl\a Pic\-

'ul.\ th:,1. a\i\t in lirc lliiiural e ill irtlnttlelit. itlcludrng PrL-ssure

li\,rr llrotoroiln ltrr'(lator\ sllcit lls ciliaie.. lilgellates. alld ail)oc-

bec \lie rolrlLbitlLls ntar ht' eithci livillg rlr inert. i'r'''lrr' ''

I-ir ing Ilicrohabitats. Spccialize-t1 lir iirg nricrol'tlbitats inclr-rde

tlrc sLrrlace'o1 lt s.-crl. I ro(r1. or lt ltal. r',hicll. uith thcir highel nu-

ir icnr 1lu.r rete und thc chlittct iiri in jtial colollizlttion b1'the added

nricr,rorgltnistlts..:ltt I)r{)tcct tllc lldclc'd tllicl'otrt'tl'oln tht'fierc.:a()nrldtiti\ c condiliotls itt thc lllliural ctlr itt,lltlle tit. Eranlplcs ill-ilrrde tht- rtse rrf R/rt:r,lrittrrt liittl lJtir'illrt' iltrttirtgicn.si.t. In ol-der trr

rnsurc thii{ Rhi-t,!tirrnt is itl clo-qe asstlciatirln \\'itll the legtttllc.

scc-rli ltrc co:Lttcl u itlt tltc rtticrtlht Llsills an rlii-t'rrgltrlislll llli\ttll'c.r.tr lllti:rtltiturt is pilrcecl irt tr brttd ulltler tlte 'ccd $'here the ne u iv

tleveloping prinrarr rool rl ill ilalletrlltL'. Itt conlrast' llttcillLt.s

rltut irtt,itrt.sis (flTt i: piaced on the sull'ece ()f the plallt leal'. tlr thc

p1:int is.ncinecrc(i to cottteirt tile tll- gcncs that allou'the plodtrc-

tion of the loric pt'otein in sitl-1. once rt is in-sested. Alier ingestion

br tlic targcl orsanisrtt. tite toxic protern u'il1 be ri'ithin the diges-

ti\. tlrct rrhele it is tttost cllectile. ir ':" '' ', \':, tr i " :ii,,.i.!'iiLIl\ ii:ril I ;l

Inert Nlicrohabitats. Rccentl,t' it ha,s been tbund that nlicroor-ganisrns can be added to natural communities together with pro-

tective inert n'rrcrohabitatslAs an exanlple. if microbes are added

to a soil uilh rnicroporous glass. the sun'il'al o1'added microor-

Sanisnrs can be ntarkedlv erthanced. Other microbes har"e been

t,t.:err etl t() e rartc their ou n ttlicrohabitatsl N'licrrrtlrganislls in

42.24

What factors m;ght iimit the

addition to a soil or water, toability of microcrganisms, alter

be able to Persisr and carry out

Chapter 42 lndustrial Microbiology and Biotechnology

*,ater column orerlying PCB-contarninaied sand-clay' soils

rbeenobsen'edtocreatetheirou'n"clavhtrtches"bl'bindin-es to their outer surface s rvith extrpolysaccharides'

'l'l-resc il-

ations shorv that *ith the application o1'principles ol t-nicro-

eccll0gt, it nla1., be possiblc tO iI()re succcssl.u]il, I.llllnAge

robial cotlli-t'lunitie s in neture'

processes such as changes in bioiilm concentlation gradients. It

is possihle to it-]easure the concentration ol sttbstances from

nranv diffcrent environntents (table 42.13) Aoplications in-

clucle rhe detectiOn ol' ciuci)se. acetic acld. glutarnic acid.

ethaltol. and biochcntical orrsen d'-:tnand In addition. the ap-

plicarion ol biOsensttr.s t() rrer\Lll'e cephaloslr,rrtn. nicoi:rliu

lctil. lind scr,e i'lil B r iilr1liirs hlis bet'n de scrrbed. Ilccentl'v

biosel.;ors har e he en i131'elirpc.1 using inlilluilochenl:ca1-based

detectii)n s\st.ir\. l-htst ner'r biirsensors riill deteci pathog'ns'

herbicides. iorills. pr()tcillS. alrd DNA. \,1anr' of the se biosen.

sors lr. blised on the use o1'a streptavidin birrtln rL-eotnitir)n

svste1ll tTechniques & Applications 42''l)'

i?.epid edr tllaL-s lLre heing nta.lc ln al1 iLre as tlf biosen-qrlr tech-

nologr. The 'c lne 1Ll.1e llrir.lili lllrfril \ clllL.iil\ irl the siabilit)' rnd

.lrr"iril,n o1'lltcse units. r,, iiich are he illg nlaiie tllttre portable attd

sensitir e . \1i.rL,r:rgltlttsttl: allri rtlctllIroljles such ls glucrlsr: can

be ntcttsrtrc.l. ihLr: rllec-ting clititlal ngg{-s irl tllorlern medtcine'

M icrcarraYs

.\ lurt.' L]i11i ,ii tl'r. llt-\r :tllrl dcr ilitllirlg rrlie i obial biole lhn0lrrs-r

tl. l.' 't I)\ \ '(rirl(ll.(\ lll gt'n('llrril\\ lr'lllr'Iiliilt

!-llc t\|1f :'1|il ill r'()llll)]c\ biilltrllclll \\:i1'lll\ tr"'ir'r'llrrlt 1-! (;)

il,. ,"1..'.i iiLl\ il;rLL,. tlrer hltr d r.)r.cL1r1.r,ri lir thi, lire ir ai.L- tha re \uit

(r1 .r \ r!le \\ itt g.-:l,,tlliu.. i'atlol]lirilliilll I)\-\ tt'clln' r1l-'r ' optiur'

Itrtr,i;1.'\.-'rl,r'.irl.riill:i1']'' ''i:'t:'lr't1rrttii"11''I;i|;"'qt'

ing. fhis nticroarrav tecltnologv ila' hccll srtgucslctl to proVici:

thi cquir elL-nt (r1 lhc cilcrlli\t's pcriodic tlrbie lt ol1ers the potetl

trtl ol lis:ltr ills .ili gt-rlcs ttsttl ltl essellli-'le atl ilrganisnl anci cltn

nt,rttit0l crp|r:ssiiltl tll'tcns il1 tltrrLt:ands tl1'gt-'l.ies based tln lhc

pr.incrple:'l,,,.,,, i,, figtrre {2.1 1' ln tilir tt-chnitlrrc' 1 0() to l00 pl

,,,1t,,11e.. cLrtlt:iillillg tlt'sllt'ti \e(lur'il'L-\- llr' sP()l1ed tlnto glltts

-t]r.jt:rli!ltIld]j]lerttllltrct.iltlslrtitltlt.irlri.l.hes.:ltrtlitsarethetltllirlt-l.L l, it}l .|)\'\: 1]ll]11 JtIl( i,\ll|.t.\\](lll rr, r,il. .i/.i1- Birldirlg

o1' tlrt .l)\.\ lrrl \ llliirll\ -gctle: it tlrta'tircri trsine rlpitl photo-

l,r(lll! l;r,'lrit"l ll,: I(i lllllLll'(''

( irtttlltlciltl lllle r()iii-la\ ptLrdltc''t IIIL- ll()"\ ar ailable that con-

tair (t.iJJ iillcll lf.ltllt':1r)l 'LIUL'llllls gcllc c\Prt's-\itllt in '!rrr't'ira-

i ()ii 7\( r,\ ( (,r.(,\ /\i.r(,. F<tr [-- t'ttlr.1.l9(t6pg1 relldrlg llatlrcs ctn Lle

\alillllcLi itl li tltl.toiift.l\ 1Ul ill'11' l(lllt''cllilll'l 'he etltire 1-- r'o1r

desired iunctions?

What types of microhabitats can be used with microorEanisms

when they are added to a complex natural environmenll-

赫hy are`Iants inoculated With θaθア/′υf_1カ

」月βσ′θβSS?

t5 BIOTECHNOLOGICAL APPLICAT!ONS7:roorlianisttts and parls t't ttlicr"()l'Jlrni\lll\' c:pecialll cn-

lcs. ul-L. rrscil rp a ri ide r ariet\ 91'hjote chtl'rlo"1.xl illrpli.ilti()lr5

l1()nitorthe]erelsol.CfitiCalct,rllptlrtlit].itl1i.tc.:ilrl]()Illll!lll, in rrtirttal,s and hullllills. These tccilllitlLtc' hltr c rr ldc a1l1t'1iclr'

ts iJt el]\'iroillllentlrl scicncc. .inirtlai lltttl ltitlttllt.i ilr',tltll. .ltrJ it-'

ic science.

)5ensor5-apidll, tlevelopiilg llr.ll of hiiltechnolocl' 1i111gti t.ll i tltetrsr'

srnational scit'nlif ic irtterest. is that tl1'biosensor proriitctioll'

this lieltl o1' bioclectrorlics' livillg lllicr()organi\ttls l or the ;l

,\'1lres ol orgatleiics) are ltnked s iiil ciecilirtlcs' atld l-'i1rlo:-l

re actiotts ilre converted into clectlicel cttrrttll' bi tllr-'c

)scrrsors (figure 12-20) Biose nsors a|e ltcitlS ilcr c-lopttl t''

rasr.ire sllecilic coltlptlt'tctlts in bt'ct lI ltlIllil' rt POllLtialllt' 1Lr

icct f.iari.r- cr-.r.tp.u'cls in i'o9ii. a1d lo :tur-ir, r'r'rr irirlttllciltlrl

Molecule discriminatingfunction

誓逮癬（ＳｕｂｓｔａｎＣｅ¨ｍｅａｓｕｒｅｄ

E lec t ric

sig nal Biosensors: Potentialand Environmental A

Biomedical, lndustrial,ications

Clinical diagrrosis and biomedical monitoring

,A.gricultura)- honicultural. r'eterinalv analysis

Derection of poliution. and microbial coniamination ot uater

Fermentation :inalr sis and ctrnlrol

\'frniarrino ,rf inrJrr<rrirl oere< 'rnrl lirrrrirtt

RecePtor

gure 42.20 Biosenstlr Design' Bitlseils'rr' 11q- i'rr'l'tr.: rr'-

r:asine applicatiotts in Inedicine' inductrial 1111;1-irlrllrlrr'gr ' riilJ

Receptacle substances(enzyme, antibiotic.

Physical and

chemical change

〓一

J…Bi5tin Bindi and Biotech

Egt u'hite contains man-v proteins and glycoproteins u'ith unitlue prop-

ertie s. (Jne of the most rntcresting. w'hicn binds tcnaciou-sl)' to biotin. *as

isolated in 1 963. This glycoprotein. called avrdin due to its "ar id" brnd-

ing of biotin. *as suggested to pla-v- an tntportant role: making egg rlhitc

antimicrr)bial hy "ryirg up'' the iriotin neeilcd br'rlanv rr,i!r, u,rg'rttisitl:

Ar,idin. uhich fiinctions best under alkaline coniiitiot.ts. has the hishcst

kno',r'n hinding utrnrtl betueen a proteitt anii a lrgard. Scveral Iearslater. scientists at I'iclck & (-o.. Inc. discrrvet'ed n sirttilar protein pro

Cuced b1 an aclinonrlccit. SlrrTr,orini a.s uvirlirti. nhich binds bjotrn at a

neutral pH arrd u hrch dtv:s noi contain carbth)'dri1tes. Thcsc characler-

i,stic> nrakr: \treptavidin'rn ideal biniiing acenl li)rbi()tin- antl it has'r':en

T,arget : Binder

used in an almost unlimited range of applications. as shown in the Box

figure. The streptavidin protein is joined to a probe. W-hen a sample is in-

cubated with the biotinl,lated binder the binder anaches to any available

target molecules. The presence ard location of target molecules can be

determined bv treatinc the sample u'ith a streptavidin probe because the

streptavidin binds to the bitxin on the biotinylated binder. and the probe

is then visuaiized. This derection s)'stem is bein-e etnployed in a wide va-

rietv ol biotechnoic,rgical r:pplicatiorts. including use as a noffad;oactive

probe in hy'briCization studies anc as a cntical component in Diosensors

tbr a u ide range of environi,,cnl:rl ntonitoring and clinical applications.

Not had tbr a protern iionr a "sintple" filamentous bacteriun-r!

Probes

EnzymesRadiolabelsFluorescent agentsChemiluminescent agentsChromophoresHeavy metalsColloidal goldFerritinHemocvaninPhagesMacromcLecular carriersLiposomesSolid suppofts

ｌ

ι

ｒ

Ｊ

AntigensAntibodies

LectinsGlyccconjugates

EnzymesReobptors

nanspod proteinsHydrophobic siies

MembranesNucleic acids. genes

Phages. viruses, bacteria.s,,bcellular organelles. cells.

tissues. whoie organisms

Strept:r\ idin-lliotin BindingSlstems are Finding \1 idespreadApplications in Biotechnologr.\Iedicint., and En\ inrnntentalStudies. lliich IrolccLtie o1- strr-p

iar itlrn. r pr'()tcin de r ir ed il-ottt atr

rlclin()nt\ cctc. irl: lolrr site s brq hich it crn hirld tcnlciousl\ tr) hi-i)till lnolcrl jir lcrir. 13r.rtlaelrilg Ibinrlcr to thc biotin. and a proht-.

such as a f-lLlr)rescent nrolecule. to

tire streptar idin. the target rnolccLlle

can be detected at lo\\' concentra-

tions. Target brndcrs. probes. and

applications are il()ted.

AntibodresAntigensGlycoconjugaiesLecti nsSubstrates, cofactors, inhibitors, etcHormones. effectors, toxins, etc.Vitamins. amino acids, sugars. etc.Lipids. fatty ac,dsLiposomesDNA/FNA probes

All cf the above

―― 乙亀 Z

‰j靴ゴ°百1‖][黒

AP、〈傘oい

°9

∫∫ξＣＯ「０」ＯＱｏ∽

一一０い

ごヽ■ｏ一ＣＯ夕ゝｏ

ゝ＞ｏＦ

T・ 1ご碗IヽL

ξ

∫ で

NSilllIIll[iog町)hy

Reference

Roboticprinting

Computeranalysis

rrit.ie.'fhcst al)ltiotlclle\. brrtil llou lind ill thc 1'ulttrc. tllltkc i1

sihle 1o iirllou lhc t\llrcssi()n ()f thoLlsand\ o1 genes and stuti\

blil rcgLrlatiort ()l' Itticr()biai giru th xl)d resl)onses tu ettr irtrtr

rtei cl'rang.-s.

What are biosensors and how do they detect substances?

What areas are biosensors being used in to assist in chemical

and biological m0nitoring efforts?

Describe streptavidin-biotin systems and how they work. Why

is this technique impoftant?

What is a gene array? What basic techniques are used in this

new procedure?

)pesticides

:re has been a lLlng-tenn interesi in the use olbacteria. fungi.

viruse-s as bioinsecticides and biopesticides (tatrle 42.14t

:se arc dr'firtcil as biological agerlts. such as bacteria. lungi.

t\c\. or thr-ir contponcnts. \\hich can bc used to kill a suscep

e insect. In this stction. ntajor ttses tlf bacteria. lungi. and

rses lo control populations ol.insects rvill be discussed

Reversetranscription

Bacteria

13を lcicritli tiぎ c111｀ incltlに 1ご [1ヽ :llict卜 lli β(′ (′ //``、 1ヽヽせせlC｀ ‐

1)「i1llti「 ll)

β ′/:ll′'71.ゞ

た''7ヽ お (ヽてく'/, マ/2' 1~1li、 bをlctcriulll l、 olll卜 cヽtlkl卜 lo、 ic

t()i11、 cctヽ そ1、 a、 cgctaliヽ c ccll llut cltiriilビ 、Pて

r`ultlli(l13 itiヽ 1()(luCCヽ

al〕 1lntiticclltllar i)1()tCi11モ ぃ、11l cl)、 tal thC Ptlla｀ P01‐ tll卜 :`〉 d)、 that cain

tlct a、 [il、 liCrOhitll il]、 ccticitic F()l 、PCCilic i11、 cct gr■ tip、

1:hC r(lI(1ヽlついrtiI Cr)、 itil altcl c、

1つく)｀ t'rC tO■ lkを lli:le tt)nditi():1、 in

thc llillti=tlt ilttピ l〕 lCI]1ヽ 1● rclcti、 c thcl,「 (ヽ 1い i` ll へitcr tlli、 「cを lCtヽ ヽヽit11

〔l Pl()tC tiヽ C Cnス トnlc tlnc ticti、 ct(ヽ 、ill i、 Prて (ヽlticcd(flglire 42.22, Si、

of thc ticti、 ct()ヽ in tlllit、 intcゞ I alc i1llぃ thC Pla、 11〕 ょ11lC nl卜ranc(lig―

urc 12 22わ ()(ol(■‐nl a hc、 ti=(ヽ nal―、|]をlPc(11)olc tiュ 1● tiピ h thC nlidgLlt

cel;、 a、 、11(ぃ 、、Il in llgtlrc■ 222て / Thi、 lcnd、 lo the l● 、、く,foヽ nl、 )tic‐

balallcc alld Al｀ Il(11ld inall)lo ccH l)i、 1、

Thc nlost rcccllt adヽlallccs in Our ulldcrstandillg tDfお`7(′

〃ιィ∫

′′7“ ′マ′′gノど,7,′ hゞa、 c in、 ol、icci tllc crcation of pest― rcsistant plant、

Thc llrst stcp in thisヽ olkヽvas to inscl■ thC tOヽ 11l gcnc l■ to f (こザ′

This、 ork sh()ヽ cヽd thal tllc c「卜、ttll IDl(■ cin cotlld bc c、 Prc、 、Cd in

tli30thCr()rgarli、 111[1,ld that thc to、 in、、(1、 cffcctil c Thiヽ ■■110r

、ciclltii｀ ic tidヽ tl I]CC ｀`tlヽ 1()llい ヽヽcd i11 19 「ヽ 卜) lhc p:.｀ tit:cti、 I]()f

tonlato plant、 thatご (ヽ IItiincci t11ビ tい 、11l gcllc

B r/7″ ′

'′

7ド た'′ :「ヽヽ ctin bcピ 1(,ヽ 1ヽl il〕 ic「 1lncr]tcrヽ ｀`卜ごFi tilc ccll、

1卜 、e thc sp()rcs tllld Ciゝ 、(al、 ctrc rclctl、 c(l illt()thc incdiunl Thc

Chapter 42 lndustrial Microbiology and Biotechnology 42.26

Test 師ヽ…∞メ凛tミ 遷 纂

奪 たPCB anrplificationpunJ;cation

3

亀:息 :1‐ 3?〔

:,=R=I=二 i

Tt:e Use ofOlder Tech

Bacteria, Viruses, and Fungi As Bioinsecticides: Anwith New Applications

Bactena

Viru-ses

Fungi

Bacillus thuringiensis and BaciLlus popilLiae are the two rna-jor hactena of interest. Bacrllus llitlrirlgietlsls is used

on a s,ide varietv of vegetable and field crops. fiuits. shade trees. and ornamentals. B. popilliae is used primarily against

Japanese beetle lal,ae. Both bacteria are considered harmless io hunrans. Pseudomonas.fluorescetts. u'hich conta;ns the

toxin-producing gene fiorn B. thuringiensis. is used on nlaize io suPPress black cutrvorms.

Three major Yirus lroups tliat do nol appear to replicaie in uirrin blooded aniinals are used: nucleil polyhedrosis l'irus(NPV). granulosis virr.rs (GV). and cr,toplasmic polvhedrosis virus ICPV). These occluded viruses are more protected

in the en\,ironnient.

Or er 500 iiifferent fungi are associated s,ith insecis. lnfcction an,J diselrse occur prirrarilt' through the inseci crtlicle.

Forrrnra_1or9ene., lr"lgheenused.Bar:rr,i,ria btssiattotuttllletarhi:iuntlni.top!iac areusedfbrcontrol of theColoradOpotatobeetle and rhe froghopper ir -su-sarcane piartations. respecliveiv. \;erriciiliun leL:ttnii antl Entontophrhora spp., have been

associateci rvith contrrtl ttf lphids in greenhouse and t-ield environmenls. Coelonrtr:es,:i chytrid. also rs beirrg u:ed ibr control ofniosquitoes.

Toxin binding to

phospho pids and

i∩ seniOn intO membrane

↓NH2 COOH

crystal

Plasmamembrane

Alkaline outcontents

Y

了 躊 鸞 り

/露 獅

側 腕 耐

◇了了鸞扮Protease

+68 l.Da oct're to.ir

--t- ..A.-/ sH ici

t\-t1-

SH

inside ce‖H20,catons

|

iigure 42.22 'Ihe f Iode of -.\ction of the Bat'illus thurinsietr.si.s Torin. (ar Rcleas. o1 the prot()\in fronr the parasprlral btldv and

,iell e: il lr)ss (ri ATP. resultrnc in ceil irttbalrrnce and lrsrs.949

Aggregation andpore formation

H.O cations

ノ

Inside cell

lnside ce‖

Toxin protein Efflux of ATP

ion channel

Outside cell

Microbial Groun lHaior Organisms and Applications

42.28

990 Chapter 42 1ndtlstrial'ν4icrobiO10gy and BiOtechno10gy

p°

電it胤:曜よ」:∵習滞llsPθttltlc｀ uscd Ю COmbtt tC

思模「蹴ittll薫IW[1聯lW撤猛瀑}‖

躍Ⅷittti:嘲 1∬漱1∬]i小鋤dと、

Viruses

鮒鮒III(llttlulぎ :ヽ(1撃ボ∬lCPVヽ alC tinc lCa、 t hOSt Spccillc tiltlscS,[｀

ffccting(1卜 out 200 dil｀

fcrcllt typcs()f inscctヽ ´ヽ1111■11)01ttllll C0111lnCrCiを lヽ 1ヽ1(li pCヽ tiCidC

i、 11larkctCd tllldCr thC trを ldC llalnlc Elcal fOr co11(rol ol｀t1lc cOtt011

b()11ヽVOrnl〃て'″`,r力

′、こ`て

′

(Dnc ol｀ thC ll10ヽ l Cヽ Cll■ 1=〔 1(1｀ そl:lCCヽ 11〕 、ol、 しヽ thc tlヽ C ●1 いtic―

t1lo、:iru、 cヽ thを,t htlヽ c bCCll gcllビ 〔iCtlllヽ 1110dilic(110 PI('(lt,cc ti lつ

て'lCnt

淋 ‖ ギiltttだ ]罵 illll懺il書li繊 |ギ麟 illl

thc l cCO11lbillを lnt bttctll()、 iltlヽ Prc

t()、 ill it tictヽ 1110rC rapi(11ゝ 'thtlll thC l)tl lじllt｀ lrtlヽ 、a11(1 letli tit111■ 1=Clぃ

inscctヽ 1ヽ lllarkcdly dCC,Cascd (|・‐1・ ■11、 ■iヽ ttl ■■t｀ 11 ・

‐ 1

Fungi

Funtiホ o Can bC uscd Ю COnは'in、

cCt PC｀ ｀「

ungalい 1● in、 cdl―

ci(lc、 をlS ll、 lcd ill tablC 42 11、 arC i′ ilndillg lncrCa、 111=tlヽ Cl]n tl=ll~

劇 Imで nedcヽdOpmo■ o「 boPcゞ にdC、 ム PO,C｀ い1腱Ⅲ

dゝ

Aヽ そlilalぅ lc biOill、 cctiCidCs Vヽhich ttrC tlCriヽ Cd llヽ'111 ltingl l11-

cludc ktlヽ tlganlyCillを 11ヽ (lthC l)ol、 0ヽ 11]ヽ t in titlClitioll 1ヽ)CCiを1l111lCro―

bio10=iCJ nndaboliに ヽヽuCh a、 hikkOm、cin and由 CヽⅢnぃ、卜n、 mで

actlVC agaHlst il〕 SCCt｀

The use of nlicroorgall56l-s itl industrial nlicrobiologl' anti

biotechnolo'r,. as rltscussed irl this chaptcr' does ntlt take place in

air elhical ltnd ecological r acuutlt Decisitlils to illlrke a parlicular

product. ,,,,.:1 tilso thc nlethtxjs used' ciin hale iong-teil"-',11:l-

i.,-t -un.r.p..rerj

elTects' as u'ith the apllerir-ance Llf eniiblotlc-

rcsistattl Pathogells lrountl the rr orlcl'

It{rclol.'io}ogv is a ciltical p'irr' of the area of indu:trial ecologl '

concenred *irh trackrng the tlor^'' of elL'nlt'nts atld col'tlpouncis

thoLrgh tht nirturili an't slcial r"orlds' or thc bii;sphere lirld ihe an-

t.l.rrosphcre. \licrobiologr" e-'peciallv ll\ an rtPplied discipline'

siroLrlrl bc .{)rl:irlere(l " itli'l itt stlpporting tt,g111l i"-trrld , :.-

\1iat ()r)rg"illisttls liave bce ll of itnt-ne ll-se berlel-it to llutl-tlttlttr

rl.iri,Lrgh rhc-ir rolc in riiorl prodLrction anj proccssilt 1l: :):l"'

their ir,,.iLrctt to itllpror e hLt'r'lrn lilil anirllal health' in iisrlc''rlttire'

artrl l'Lrt ril.' Iltainte nlttlct- ltrltl itltprilrelnettt tlf ellr ironntcrltal qtlal-

irr. ()tit.'r lttrarr)(rl!ii1ll\lll\' il(i\"c"L'r' ai'c ittiptltlltrlt pathilgetls llrltl

:..i.;: ,;, .1.,',i,,g.' 't'tt' rtlicr.t'r.krgisls hrr'e i.icipcct colltt'irl .r linlit

th.' .teti" liii> ol thc>c- ilartlrliLl tlltctiltlrganlstls''iht {i5i'ilra'1 r lnrl

rr.r. rrl hitri'iiitltl tlllcrilhrlii 1lr'tducl:' sltell as illllihi()iics' hlirc- cotl

trjl.irtcti t,, r, .lciribltlls ol thc hlrtllatl lil'e :lllirl in tile lasl 'clllltl'-\"

..\ ttttct,,lrtLrlrlgtrl ullt' 'iilt-ks ill lttit tlt tht:'' ltrelLs tll'brtl"t'cil-

ilrll(l!\ \iltrrll'1 'tltl:itlt'l 1ll" i'rn'lel tt-f'lt "'tltt'ttt o1'potsiilic tech-

nicll .ieir:i'.ttls"{n trccllcrlt lllil()(itl'tl(ln t() lllc lelationship

he t*ettt tc.lrn{)l{rg} arltl ptrs:ihle sociciltl trl1plicts it Sit.t:'.]t.-'

Satrtr.tcl Fltlrtlllttl t.st't'ttltitr:ntlit l'r' ()Lir-f irst challcrlge' a\ llllcl()-

[]qrl1rg111r. is to titldcr"""t1' tt-t lllLtch lts is ptrssible' the potentiltl

lnlllxtl; ,r1'tlar" lltoiiLl't '\ illl(l P1()ce\\'\ l'rll thc Ittrr'lticl 'r'tit't\ 't:

rre l1;i: \rn iitiar()bi(ril,gt' .i" e ssclllial plll-t oi Lilrs re sprrllsibiliir is

1o h.- .rble ,,' att""t'u'ii*tltte cl'fecttr clrl n ith ihe r ltriilris 'socictll

stekelrrrirrir:.. ltirrr11r the iararetriric urtl rr,rgcr'-tc* rrotc-lrtiril ittl-

f ,,.,. ,,t rrlicrtrbirrl-baseil (rillti trthet ) tcchlltllogie t'

lMPAσS OF ArliCROBIALBIOTECHNOLOGY

讚 mm“

“

ぉ bd∬ edd“ゞ

Briefly describe hOヽヘノthe θaθ 〃′」S r力υr′βθ′θ17S'S toXin klllS

are being used to attempt to conirol in

name for one of these Products?

・踪『謡紫[11=£撃[¶事部照lroadest possible impacts in your discussion of this

Define industrial ecologv'

What are the biosphere and anthrosphere? Why might you

i古 1薫 tiビ term antlЮsphere was COlned?

a particular

terms of the

problem.1

2

3

4.

insects.

What tYPes '

sects? What

Which fungi

of viruses

is a tradepresentlY

2

3

are being used as broPesticides?

42.1 Choosing Microorganisms for lndustrial

MicrobiologY and BiotechnologY

a. Industnal nlicrobioiogr tt;ts hcen LlrL'!i lril ill:ii1Ll

facture such products as anlihirrtl" irllu:1!r

acids. and organic :rcids and has had Illlrl\ ln'l

pc,nunt p,'trtite eflects cltt anin.ial allrl hurllrin

healrh. \Ii'st u'ork in this area has btt-n currietl

out using micrtrorgantrins i'olaled tlom n:llui'

or nlo,iitied hr the u:e rri e lus'il lnutation l"h

niques Brotechnologr inr'rl''e: ihe use of mo-

lecular techoiques rtl trt'ditr lnd imprtlr:

m lL I oUrg -lni \ll I \

h, Finding neu nlicrtrorsanisms in nature lbr use in

lio,".linolngl is a continuing challenge For

nost en\ ironlnalli\. onl\ a ren small part ol th

rrbser' rhle mtcrrlt'ial irrrnmunitr has teen grou'

rtables J2-1 lnd J2'lr' bur malor adranc'-.s t

trtru ing "uniuiiuretl nlitrot'e: are bting nlade

a. i"leation an,l mulatltrn crrntinue to L'e imgrnar

approaches 1-or identi f1 ing nerl m icroorgantsm

Ttrese o etl -.stahlisheil procedures are now bein

4229

conrplertlenlc.l br nloleculltr tcchniqLres. inclLrd

rng ntetabrtlic ettgincet ittg and conlbinatorirl bi

ologl. \\'ith contbinatori:il biologl ltable'12.31.

ir i-. possible 1o tr.in\iet gL'rlr's frolll r)ne t)rganislll

to rlnother oiganism. lnd to lilrnl ne\\'pr()ducts

Li. Sile-directed tlluta!enesis artd prrotein etlgineer

in! arc uscri to rttodill gctte erprr'ssiirn. Tilese

approrrthcs arc' lcadirlg lir lla\\ .tlld L;l-tcn diller-

enl plo.lLtcts riitlt tleir Propcltics l[igure {2.3t'

c. Nalural gartelic (-n-sin.!-r.lnc. is ol inctertsing in-

r.ras1. Thi! i;tr0lres erplCitilll. rlltcrobial re-

ip()n-\c\ l0 slrcrr in lldaptirc nlutitlion arld torccii

!'\olutioir. uith tltr: hope ol idtntilr ing Itticloor

t.!r j.lll \\ rllr L r ir,'t cttr, .

42.2 Microorganism Growth in ControlledE nvironments

Ir \licro,rtgalirmr elitt i.c lrirl n in eonllrl]ltd enr t

1\)nnltnl\ o1 r llitlus t,, pc: :sin-L lirlllglli.:rs lttltl

rrlhtr eLtilttrc' \)'lL'Jr\ ll L['llncr] ctrtlslilttcnts ltle

u.cd. !r()\\th pillllllL'trls alrn be rh(lscll rnd \r jc-d

II tht crrur.e o1'Ir(r\\jllr lt Illicro(llgrttisnl. Thi'

:ririrrr)irail ir Lrt,:ri l,erlrclrllLtlJ lirl thc i-tlllti.liorl rll'

trl,,. l ',:' .r':l rl I:)r-l i'

42.3 Major Products of lndustrialMicrobiologY

:r. \ r iLlt \.ttitl\ ('i..111Ptlttrlrls rle lll'lrlllc.:iJ itl rn-

tjLt.ttirLl rttierohr(rl\)g\ lllllt itllllltcl llttr Iirr'. itl

nrirlr\ \\il\ \ ltable {2'9). Thcrc irriiutlt ltltlbr

,'iL\. alllill(j ll!l!l\. olI:llli. lci(l\. hi(rlloL\rll'l\

\ri.h J\ iitc crelLrrlcrtlitt:. rllLi hio\lilli!aLlllts

rtigures "I2.8. -12.9. rrld J:.1 1) \1icr()olglllll5lll\

rlro crn bc tlscd 3\ hi(rcJlill\\ts t1l cilll\ oilt sl)c

c j1 ic chcttt tcrl 1 !'i].iilrrls r ti gure tl2. I 2 I

,q ;. "1.;;,11'.i,.' .,'1'11', 11r,1 . ,t itt'

Irlr'1ltnl jlilll,li illrlustii:tl 111j;1o[]lrltlgV atld

biotcchi〕 ol●

=ゝ

' ThCSC incltidcヽ ●idcl)' tl、 cd anti―

ttllnor agcnts(table 42.11)

42 4 Microbial Growth in Complex Natural

Environments

a 4ヽicr()orgalli、 11l gro、,tll iI]colnPIC｀ 1](ltural cn、 i

l● ni〕 ,ciltS StiCh tiヽ oヽil、 ti:l llヽ l atClヽ iヽ 11()t tlヽ Cd t()

ciCtiC n3i(｀ l()bial Pro(ltlct、 卜tli to ctlil).Out crlヽ :

「()ni]〕 cntal l)〕 arlagc]〕 ]cnl prOcc、 、cヽ llnCitltlin=

bi()rcnlcdiation Pl`int inoculatioll and()thcl lc―

1(ltcd t,cti、 itic、 11〕 thcsc ctl、 cS tilC nlicrobc、

thclnl、 clヽ.c、 :liC 11()ti'il)aI P!(,dtict、

b Bi(,dCg「 ad(ltio11:s a clitictll Part t)fl■ lttH al、 ゝs―

tcnl、 nlcdiatcd lt、「=cly t,)nlicl o●

「=arli｀

11ls Thi、

C til]inヽ 01、 c nlino「 cha11=c｀ i11を11■ )ヽICCulc rra=´

Incnttltion or nlincializatit,n trlgurc 42 13)

c Bi()dcgratlation ctin bc inl]ticllccd b).inl a n、 i tiC―

t()iヽ inClし 1(111lg o、 )=cl1 1)iCSCl]CC()l ・lい 、cincc htl―

11lic aci(1、 alld ilnc p:c、 clヽ CC 01' I ctiて 1ll) t、 tiblc

organic illattcI Rctlutti、 c dc1lalぃまcnati()ll Pl()

cccは、 hc、 t tlllticl all(tci olpi(: conditi()il、 allti thc

PrC｀ Cncc● 1● l=aniC lユ 1[ittcr Ctlil facilitatc rll()て li「 i―

ct,11()11ぃ |「CCJc tlュ ぅtC():う ,P(ヽ 1l l(1、 il¬ lt lう rぃ CC｀ ｀

ぃi cぃ 1]lcttibい iヽ l il

d Thcヽ tructurc t,1ぃ「fanitこ )`nlP()und、 inllucncc、

dc gitt(ltiti()1]11ビ Ⅲl、 titticiill コIC ilヽ 1)ccilic l● cti―

11)i〕 、()n a 111い 1ビ culc ti、 in lhc′ 11(■ 11〕 )｀ 11()Il l fl=―

1lre 42 141()i il 、alic(1 、1l titttiI[1: :ヽ 1 1llci、 al C

PiCSClう 1 こIc=I atiatiⅢ l, caiュ 1っ c t,「itご :せ て1

C L)C=]tidtiti(ヽ ぃ |]■ l n tl=C■ c ii ctil' 卜c t arlictiい tlt i11

1,lacc 入ヽl〕 cthci tl]、 卜ch=c nl:「 1:1ビ ぃ|、1ぅ

i11、

、ぃil、 o「 l1lc 、ub、u;iatc tflgure 42.17, Such

itil ic、 ctllC Cll)11ヽ 日 tヽittlヽ 1'1ヽ いh C tilcぃ 、こぃ1「うtil―

tiral l,licr(、 bial co11lnl111litic、

「 IDc,「 adatiol]can lc(ld to lll(lca、 cd t()ヽ icil) i11

1]lal]、 ctlSCS Ii l)●t nlal13=cd cを li Cl'tl‖ ) 入ヽidc―

ヽ'ICtld P。

lltlti● :i ctin いCCtll lroll c()「 I()ヽ 1()n i、 a

Questions for Thought and Review 991

particular conccrn、 vith mcthanogcns and Dど ゞ“′―

力1'jわ ″jο playing important Юlcs in this proccss

g Plants can bc uscd tO Simulatc biodcgradation

proces“ S during phメ oFCmCdiation This Can in―

volvc cxtraction. fIItcHng. stabilization. and

、lolatilization oF pollutants([gure 42.18 and

table 42.12)

h 4ヽicroorganisllns can bc atldcd to cnvironincnts

that contain complex nlicrObial communitics

with grcatcr succcss if livitt or inCn mた roh,b

itats arc uscd ThCsc Can incitldc living plant sur―

1■ccs(sCCdS_ roots_ lCa、 cs)or incll matcrials

suで h as nlicroporous gia、s_R力`_r2み

i″″is an im―

portant cxainplc of a lllicnoo瑠aniSm addcd ttl a

COInPICX Cn、 :ronn、 cnt usin=a liヽ ing microhabi―

tat(thC plant rool)

42.5 3iotechnolo9ical ApplKations

a lヽicroorganisnls arc bcing uSCd in a、 vidc ral]gc

of biotcchnological applicatioQヽ stlth as hiく )、 cn―

sOrS(flgure 42_20) 4ヽicroarra)s arc uscd t()

monitor gcnc cxprc、 sion in conlplc、 systcllls

(tigure 42_21,

b BtlctCria 、iruscs and rungi cdll bc tlscd aS bi(う in―

、ecticide、 alnd b:()Pcsticidcs(tabIC 42.141 Bて ,(〃 ‐

/″、 7カ″′ル2マ′′″、′、 iヽ .tn i!111、 lヽtani bi()Pc、 tiCidC

and thc BT=Cnc h(卜 褥 cn inc(ヽ rp)ratcd i:lto co「 n

42.6 impacts of r゛ icrObia1 8:otechnolo9y

a 11ldt:stilai nlicIく )biolo=}and biく itcchnolo=)i cを in

lla、 c loll= tcr1ll and Po、 、ibl)uncxPCCtCd P()、 1-

ti、 c and ilcgati、 c ctl_・ ct、 く)n thC cn、 iroll1llC1lt_

and o:l allill,tilヽ and intllllan、 i:l]Pactcd b)'thc、 c

tcchnologics lヽ llヽ anccsin biotcchnolog)ヽ hOtlld

bc c()Insidcrcd in a b“ )ad cc(,lo=ical and、 ocictal

cて、intc、 1 hヽiCh i、 thC lk)で、1、 of illdtl、 t●al ccolo3).

:rrlaptir e rnut:ttiit:t 9(r9

jnlil !r\phtre 99()

hinirLrlr)entelion 9X'i

lrr()ailisi\ 51 t,)i9

hioJcr:r:id:liott ()J9

bioinsecticides 988

hropeiticide 9Ex-

biopoll rncr 977

bioprospecting 96'ibiortnrediatio;t 982

biosensor 986

hiosplrcre 990

hi0lranstoilrliilioll 97te hir al 9liO

conririnrLlor ial biologt' 966

crrmetabolisrn 981

cortlinurrtts lettl 9l:.tcr nlcntrtion 9l(.)

torced er olution 969

sene rrrt\ 986

industrial ecolog-r' 990

land farming 9E0

lvophiiization 96!rrrr,/r etlect 9ii0

iltetrhLrlic c(rltttol angin!'!'rln! q6--

rlcliib()lic lllth\\ ii\ etlg jtl.':i i illi \1['E I 96r

inieroart rr lechnologr 9s6

Irrt. .,l .rl Lrr.\",ttttJrr,'t"''inirtural atlenualion 93:nrrtural genetic etrginecrinl 969

non-Nervlr-rnian [-.rotlt 97 I

path\\,avxrchitecture 967

phrtorenrediation 9tl3

prinlxrvnielaholite 97,1

proteincn!ineering 9{.6

protopia\l lusion 96-5

rer:rrlcitrancc' 979

reductiredehalogenation 919

regulat()r\ mulanl 976

scaleup 97 I

secondarl rnelabclite 97-l

\enris\,nthetic penicillin 975

site-directed mutasene\is 966

″ ´Hor,, i i it otr:sible lo creete a niche rrr rnicrohahitat

Iilr J Inirrrx)lr'iLn,rill'\\h3{ Jre mc \pc(ul F}inltof concem in tq ing to maLi .ure the microhe can

linci its best piace tc sun ir e ard function lflqrrr miehl the prr'tscnrrmic" era difler fronr

the "senontic era'- l

l. \\-hat further tcchnologicai approacbes u i11 be

requireti to culture n.:icrcrorganistlts that have

nr)t \ et been groB n I Consider the roles of nu-

trient fluxes. comntunication molecules. and

competition.

\\'het nlikc's the rrcri tr1 n:Lutltl seilL'ila !'n!1

nr'er in{ Lrniqlre ) Isn t this :illlpir ri hrl h:: ftct:

:nl t,l , It i t ,r.rlritr .. ,.1 flr. Illlr. llri.'." -: ::

sisrr: $'cre tlrst aLrle to lLrnction l\\'hat are the advanteges o1-microart:ir: for ihe

\tudv ol sene cxpresrion in compler organislli: l

|し ″‐

中議

口Ю

Ｏ

（じ

♂―″

Ｌ　貞質Ｖ

面螂　請

:ndLIStFia:

o!ogytechnology

Concepts. lu4icroorganisms are used in inCustrial micrcbiology and

biotechnologi' to create a wide variety of pr-oducts anC toassist in maintainrng and improving the environment

. Most work In industrial microbiology has been cai'ried out

using microorganisms isolatdd fi'om nature or modifiedthrough muiations. ln biotechnology, microcrganisms withspecific genetic characteristics Lan be constructed to meet

cesrred oblectives.

" i\4ost microorganisms have not been gro'"^/n or- described A

rnator chalienge in biotechnoiogy is to be abre to grow and

cl-rara.terlze -ihese observed but uncultured microorgan-isms rn ',",,hat is called "bioprospectit'r9."

. Forcecl e';oLuiion and adaptive mutatlons now are parl ofbrotechno ogy. These .:L De carried out in processes termeC" natural genai-i- engineerlng."

. The developmen' of meciia and specif ic conditions for theqro'.vth of microorganisms is a large part of industrial mi-

crobiology and biotechnology Mi-

一一■

crocrganisnrs can be grown in

controlJed environments rrith spe-

cific ltrnrtations to nraxinrize thesynihesis of desired Products.

. Microbial gro'',r'th in soils, waters,and other environments, whereconrplex nricrobial cornmunities al

ready are present, cannot be com-pletely contrclled, and it is not possible

to preosely define ltmititrg factot's orphysical condiiions.

. Microbial gr-or,'.th in ccntrolledenvironments is exPensir''e; it is

used to synthesrze high value microbial metabo-lites and other products for use in animal and

Biodegradation often can be facilitated by

changing environmental conditions.

Polychlorinated biphenyls (PCBs) are

u'iiiespread in<justrial conianlifl ants that

accunrulate in anaerobic river nluds- Although

reductir e dechlorinatirrn occurs under these

:.rnaL-r()hic cr.rndili(rns. ()\\ !€n is rcquired to

.()mpleie the dcr:radatitlrt process. In this

erperinrenl. nruds are being acrated to allou'

the final biodegradaticrn steps 1cl occur'

一ネ一

一．一、

一一一一一一ヽ〓Ｆ・〓

ギ・ [1警ξ手

||ギ 11

′「

ｉ一．・．■一

′れ

・壼．●デ鷺

）世ξｉ

ｌで■ダ

あ と 脅 〃

96,4 Chapter 42 lndustrial Microbiology and Biotechnology

human health. ln comparison, microbial growth in naturalenvironments usually does not involve the creation of spe-cific microbial products; microorganisms are used tc carryout lower-value environmental and agriculture-relatedprocesses.ln controlled growth systems, different products are synthesized during growth and after growth is completed.Most antibiotics are produced after the completion of ac

-: .' ,. ;.tive growth.Antibioiics and other micrcbral products continue toconti-ibute to anrmal and human welfare. l.lewer prod-ucis include anticancer Crugs. Comb!ratorial bioiogy is

making ii pcssible to produce hybrio antibiotics rnrithunique properties.The prociucts of industrrai microbrology rmpact ali aspectsof our Iives. These often are bulk chemicals that are used as

fooC supplements and acidifying agents. Other productsare used as biosurfactants and emulsificr-s in a r,,ide varietycf applicat;orrsDegradation is critical for understanding rnicrobial contri-butions to natural environments. The cherriicalstructure ofsubstrates and microbial conrmunity char.-.cter strcs piay rrn-portant roles in determining the fate ol chemicals. An;ero-bic degradation processes are rnrportant for the initialmocji{icatron of nrany conrpourds, especiaily thcse r.vitlchlorine and other halogenated functions Degradatroncan produce sirnpler or modified 6,,111,r6snds th;i rnay notbe less toxii than the original compoundSirseniors are unCergcing rapid de;,eloprrent, which'rlimited only by the advances that are occurring in rrolecular biology and other areas of sr ience. lt is nor,n, possible, es-pecially with streptavidin biotin-linked systerrs, to haveessentially real-tinre detectron o{ important pathogens.Gene arrays, based on recombinant Dl'JA technoiogy, allo,,vgene expJession to be rnonitored. These systenrs are berngused in the analysis of corrplex nricrobraisystems.Barteria, fungi, and viruses are increasingly employed as

bicpesticides, thus reducing dependence on chemicalpesiicides.Application of microorganisms and their technoiogy hasbotir positive and negative aspects. Possible broader impacts of applications of industrial rnicrobioloqy and brotech-nologl, should be considered in the application of thrsrapidiy expanding area.

'l'ltt nti<'robe will lnve

ndr-rstrial microbiologr and biotechrolr)cv both in-

" olt e the Llse tll- illcr,rr)r'Slni\nr\ ii) ir.hieve spccific

gotll、 、hctincr crctiting nc、、 prぃてluご 1、 、、ith lll(ヽ llclttrヽ aヽltlc or

illlprぃ 、:1lt thc cil、 irl‐ nillci1l IIntitト モ:itll l)liご lぃ 卜:ぃ 1● ■、 ti、 11 1,ti、

:itiditi● I]till卜 dc、 clて 、pcd focと ,、 ビ、、)I]I]iい dt'こ 1、 、tiChよ ヽP11ま ::ミ ltiCC t]―

[iぜ tll こnl1 11lcdical connPoし lnd、 (ar,Iibit)liご 、 11て「ヽnl ollcヽ 1l an、 一

fo「 :rlcd stcroids, solヽ iCnts organic acids.chcnlical lbcdstock、

422

anrino:rcids. anci enzrnles tilrit harc direct economic value. Thenricroorganisnrs cnrplorcd bv indLrstry hrre been isolated frorrnalLlre. end in rrurnr clr\c\. \\crc rrrodified Lrsins classic nrulation-sclcttion pr oceclurcs.

Ilioteehnolog', hu: l Iong historl. rnvr-rlving ti.ie irsc of liling or-ulrrrisnr: 1'oi' conrnrer-ciiil hcncf-i1. In thc lasl \,'\ eral decacles. the

tL'rnr hlr\ i-lacr-rnre usstlciltted u ith tire trOcliiicltiitrn ()l' ltliLror)r

sairisms thloirgir tht- usc o1'ntoleculur hiologr. including the use

irl recornirinlLnt D\,\ Iechn()l()g\ l.:ta <lrultttr l+). It is no\\'pos-:ihic to nlinipLrllrtc r.nL-tic inf'ornlrlion rrntl rlesign Ilroducts such

as pro{e ins. or Lo nrorlilr rnicr-obial gcnc e\prcs\ion. In iicltlilion.gcnciic inl'or-rnlilion e lin be ir-rrnsicIred b.'tu e eir nrerkedlr difler-e nl !r-oup\ ril'or'!lnirnrs. r,uch lrr hcluecn llictt'r-ia anil plants.

l-irc us.: ot' nricroorglinisnrs in industriai nrir:irlbii'rlir,cr, and

biotr'e hntrlogi lollrrn s li logiclil \cLiLre nca. Ii is irecessarv first torrltntrir ()r' ! r'tlrlrt lt nticroorgllti:nr thlil clilric: out lire clesir-cti

[]r'1)LL'\\ in tire rrrosl tll'ieient rlllinrcr.l-his nric|o,rrgenisi| iiren isii\cil. cither in.i coirtlollcd r-nvir{)nnlcrrt:rich lrs rr i'ernrentrrr-oritt c,rntplcx niitullll \,\ \tL'nls. sttclt lis in srtils or \\ ll[r'rs. 10 lchiu\ e

.irtiilic Srri j..

CH00SiNG A/1iCROORGANiSMSFORINDUSTR:AL MiCROBIOLOGYAND B10TECHNOLOGY

「 llc iir、 (ta、 kl()l(tn i11(l tl、 tritti l1licl obiol()ti、 1 1、 1()llin(1:1、 Lli t(lblc

11licr()o「 tani、 11l olnC that is tc1lclictlH、 ‐ 、lti卜 lc ctt、 、 to l〕 1[lilltt、 ill

(lndゞ |● ｀` tinCiヽ、cll、 tlitc(11()l c、 (l tt(■ it)nt,1 、cIっ alを 1li、 )1〕 ()十 (lc、 1lc(1

1)|()dtiCl、 ,へ 、、iclc 、r tl「ict卜 ol tll(Crl〕 ati、 c al)l)lt)を iC1lC、 (Hct,ヽ ailabic

I[11〕 ■int fr()n、 i、 ()l■ tiini n3ic,,()(01■ tlll iヽ 111、 1'1(ヽ 11l ihC C11、 irolllllCnt t(う

u、 inぎ 、●1っIni、 tictttc(1 121()lccuLtr tccil:li(luc、 t()l11()(11「). tlll c、 iヽ ling

nnlcI()()1 2tti,lヽ 1〕 ]

Finding Lttici′ ooroattiSms tn ttatttre

tinlll rclati、 cI卜 「cccnti卜 thc 11ltl(1、 ()urcc、 、)F nlicI()bittl culturcs

lol‐ t卜 c i1l i1ldtl、 trial 11licI(♪ biol()t、 、、CrC llttttlltll 11〕 11lcl iclis、 uch tt、

、oil 、ttinlPlc、 、、latcr、 そ11ld、 P(,ilcd卜「c tl(l a11(111‐ tli((｀ tllturc、 1l ol“

aH tllca、 of thc、、orld contillしlc lo bc cヽ そinlincd lo idclltilヽ 1ヽ 11[1111ヽ

、、itll clc、 il t、 1ぅ lc charactcli、 tics llntcrc、 t in huntint f()r nc、 1111-

croolga,11、 1111、 、ol biol)rospcctillg,colltilluc、 loda)

Bccau、 c olll)a lllinor Portion()f the nlicrobial spccics csti―

n]atcd to cxist il〕 11loSt CllViI()nnlcnts has bccll is()latcd or culturcd

(table 42.1)tilclc is tl continuillg cffol■ thl()ughotlt thc、vorld to

gro、、 lincsc しlnctllttlrcd・ 11l icl()o「 gtlnis11ls 1ln tCrllls of thc llll101-

c11、 ir()nnlcnt、 11l o、 i nliClobc、 that can bc obscr、 cd ha、 .cn()t bccil

culitl「 c(1(,「 icicntilヽ c(1(table 42.2, ヽヽ:ith i11ビ

l ctlヽ CCl ill(crc、 t in llll

cIぃ いitil di、 cl、 1[〉 Iュ liご「()卜 itll cc()log) an(l c、 Pccia‖ ) i11 1lnicrooI

gani、 1ぃ 、 1l olll cヽ tl C 11lc c ilヽ i10n:1lcnt、 {Tcchniques &rゝ pplications 42.1). 1llicI()biologist、 arc lcarning hO、 lo gro、ハ「

964 Chapter 42 lndustrial Microbiology and Biotechnology

human health in comparisOn′ microbial groぃノth in naturalenvironments usua‖ y does not involve the creation Of spe―

cific microbial productsi nnicroorganisms are used tO carry

out lo、ヘノer Value environmental and a9riculture― relatedprOcesses

O in contro‖ ed groν、/th systems, diferent products are syn―

thesized during gro、へ〆th and after 9ro、へ/th iS cOnnpleted

4ヽost antibiotics are produced after the completion of ac―

tive gro、へ′th i ′ 二 ′・ Antib10tics and other naicrobial produ(ts continue to

ilよ留l:∴ 11『 :infrl」∬誂∬|:鷺 rti:ふ島ニmaking it possible to produce hybrid antibiotics ν、/ithunique properties

e The products of industrial micrρ biology impact a l aspects

of ourlives These Often are bu kと hem calsthat are used asfood supplements and acidifゞ ing agents other prOductsare used as biosurfactants and elη uls fiers in aぃノide varietyof applcat ons ヽ ′

・ De9radatiOn is crit cal for understandi∩9 nni(robial cOntri―

butions tO natural env ronrnents The chelnicalstructure of

substrates and micrOblal cOnnm_ln ty cl‐ 8raCter st cs play,fn―

pOrta∩ t rolesin detertnining the fate_lfchenn cals An3ero―

bi( de9radation prOcesses are inlportarnt for the in tial

modification of nnany colη POul:dS′ especia y thOse 、4ュ thchlo「 ∩e a∩ d other halogenated ful¬ ct Orns De9radatior¬

car]produce simpler or n¬ Od fied cゥ nI「 OundS that lTay lnOtbe less t,xic thall the ori9inal COmpOund

・ BIosc n,Ors are under9oing rapid develoll1lnent′ 、Aノ hich Fli「nited only by the advances tinat are occurr r¬ g in iη Olecu―

lar biology and other areas of science ltisinOぃy possible′ es―

pe(ia‖ y ぃノith Streptavidin― biotin inked systems′ to haveessernt a‖ y rea tinne detection Ofi:mportant pathOgens

e Gene arrays′ based on recombinant DNA techno109y′ a lo、ハy

99'C eXtrF:ユ 1911 19 be mOn,tOled These systcms are be ngused in the al‐ alys s Of cOnlplex n)icrobial systenns

・ Bacteria′ fun9i′ and viruses are irncreasin91y er〕 np10yed asbiopesticides′ thus reducing depe∩ dence Oin chenn calpesticides

・ Applicat On of mlcrOOrgan sms and tino r techno10gy hasboth pOs tive and negative aspects Possible brOader im―pacts Of app ications ofindustrial:microbiolo9y and biOtech―

1¬ olo9y shOuld be cOnsidered il‐ the app ication Of thisrapidly expanding area

422

arrrino aciils. an(l enr,\ ri)es thrit hlir r- direct econitnric l.alue. Thenricroorganisnts ernplorcd bv industn hare bcen isolated frontniltr.lie. and in nlin_r cil\as. \\ crc nrociilied using classic rnutation-srlt-'et ron procecl ures.

lJiot,-chnt;lo"r hus li long histun. lnr,c.;lr ing iite irse of lilin-e or-!iittisttts foi con.rntr'rciril hcnef-i1. In the last stvcral dccacles. thete

'nr hlrs bccr-.n)e uss.ciete d u ith tire m.rirf ication of nricroor-

qanisnts thr.ouglt the rr:e r-rl'niolecular hiolrtgy. includins the useo1 r-cconibinunt D\.\ lechnolLrgl (tt,e t ltttltrrr /4). It is no\\r pos-sjblr io nlrrriprrlute g.nelic intorntrlion rinri desi!,n products sucha\ pr(itcin\. or to tnodilr ntretlbilil gcne e\pression. in aclciitiitn.gcll.lie ittliit tttrttirtp tltp bc translc;rerj br'11ren ilarkedly clilfcr-ent grouit\ o1 orLlinisnrs. ,.uch tis hetu ce n 1:lre lcril and plants.'I-lri:,sr .i,ticr..r!u'tis,ts i, i.dLrstr.iui l,icr-,bi,Jloc_r, anclitiotte hrtrrltrli lolirin s a louicitl scqLlcnci.r. lt is ne .e:srr1 Irrst iordr'ntil'r {)r.!lairlo lr nricloor!lrnis111 111,,1 cer-r-ic: out rhe desircdllr'(rLc\\ in lhc nrosl clf icie nt illllnndr. l-his nricroorg.rnisnr iiren isu\rrrl- cilhc-r in ri r'ont|0]lccl rnVrronnrent sucir as 3 ierntenlcr 0rtttirr111|li'1 niltLiJel s,\\tclns.sLrihlsinsrtil:or\\alcrs.tlrltchicr,e.1te.if ir'Sir:rI.

CH00SiNG A/11⊂R00RGANiSA/1SFORINDUSTR:AL MiCROBIOLOGYAND BIOTECHNOLOGY

‐「 |〕 c lir、 1[を 1、 k lol(ln il〕 (i tl、 tritll illicrol)101()=i、 ( 1、 lo l｀ illdをて、tlittll)lc

ill ic「 (,o「galli、 l11 01RC thtit i、 gcllctica‖ )、 ltiblC Ca、 卜 [(D l]lai11ltlin

tinci=r(,、 `al〕 d、、cH、 tlitctl l()1 じ、〔laclion()1‐ 、cPttl 11(loll ol(lc、 1lcd

l):('CI tlCl、 :ヽ 、、1こIc 、aricl) (、 1(11(Crntlll、 c tll)lD:()を lChCヽ alc a、 1liltllDIc

ra11ビ i:1= f10111 1、 01:lti l〕 ゞ1lliCI()()1‐gani、 111、 11()111 lhc cnヽ il()[lnlcnt t()

し1、 in= 、01)lliヽ tictltcd 111()lcctlitll tcchllicluc、 10 nl()cI‖ 'ヽ all Cヽ i、 ling

nllcI()。【tanlヽ 111

Finding MicrOOrsaniSItts in ttatttre

tlntil rciati、 cI)rcccntl)lhc ml(1ヽ 凛lcビ 0`1 micRbiJ culmrc、lk:)I tl、 c in in(ltl、 モlial l〕 nicr01っ i(,loご )、 で`「C Intlltll tll lllalcl ial、 stlch aヽ

、()il、とllnIDlCヽ ヽヽla(cr、 and、 Poilcd lDlCを 1(1を 11ld ll■ li((「 tllturc、 lTonl

[Jl alcti、 (「)li thc、、01ld c01nti11し ic l()hc c、 a1llillc(itO i(lclltifv、 (1を li11、

、it11(lcsiI(1卜 lc ch(1lactcli、 tic、 Intcrc、 l in litintillЧ ll,I‐ nc、、‐1ll i

croolg:lnisi11卜 、ol bioprospectillg,cOntil〕 tl c、 10(la)

Bccatl、 cO:11)a11linor portion()f the llnicrobial specics esti―

1lnatcd to cxist in lllo、 l cnvil‐ ollnlcnts has bccn isolatcd()1‐ culturcd

(table 42.1).tllcrc is a contintlillg cllbl■ throughout tlle、 ハ「orld to

gro、、thcsc u1lcul(tll cd｀ 11lic100「 ganislll、 11l tcr11ls()f thc lll[l10r

C llヽ 110nIユ lcnt、 n]()、 l nlicrObc、 that call bc(♪ bscr、 cd lla、 .c ilol bccn

culttll cti()l iticlniliヽ c(1(tal)le 42.2, ヽヽ=ith i11ご「c tlヽ Cd intcrc、 t in ilni―

cII)1う itil di、 cr、 iぃ 11liC10卜 itll cc(、 lo=) tt11(l C、 pccia‖ 卜 i11 1〕 ]icroor―

gュ 1〕 1、 in、 「1()1〕1 ビヽ tl cnlC cn、 11‐oll11lCntS (ltchniques &

へpplications 42.1, 1〕 nicit)biOIogist、 arc lcarning ho、 lo gro、 '

′ノ

／

7功ピ777'こ ′て177ピ ″J〃 /2aν

`″ι`Za,1

ndustrial nlicrobiol()g)and hi(、 tcch:1()10ピ ゝ‐ b01h in―

L/ '.'olr e the usc of rnicroorglinisnts itr lchier e .pcciiic-!oalS. it he ther= crcliiin-q na\\ itr.oalLtLt\ \,, tth nt()netai'r, r 11111.. ,11.

improrinS ihe environment. Indu:rrial nticrobiolocr. as it hasIraditir)nallv der eloped. t-ocuses on prL)ducr\ \uch as phai-ntaceu_tical and medical compounds iantibiotics. hormones. trans-formed steroids). sol\rents. organic acids. chemical feedstocks.

423 Choosing Microorgan isms for lndustria I Microbiology and Biotechnology

ilic Archaea in Biotechnol

There is -rreat inierest jn the characteristics of archaea isoiatc-tl fronrthe outllo'rv nriring regions abovc deep hvdrothermai vents ihai re

iease rirter at 250 to 3-50"C. This rs because these hartlv organisnrscan -gro\\r at ierrpeiatuie-s as high as l2l "C. The probhnts rn sr{,u ingthese microorganisnrs in a laboratory are formii]able. Fcir examnie. tirgrou,some o1'them, it is necessan,to use spcciai culturing chanrbcrs

and other specralized equipment to rneintarn ulier rn the liquiil state

at these high temperatures.

Such rnicrrrrrrganisnrs. terrned hr,pefi hernrophiles. u ith optilnurnlrou'th temperatures of 80-C or abuve (.rr:c pp. )2-i-l).1t. eonlrLrnitunique chrlierrges in nrtrient acqursition. nrctrrbolisni. nucleic acjtirepircation. and grorvth. \.1anr oi.these a;-e inaerobls that dc-pend on

cle ntental sullir tr;- 1in ic ion as oridants. Enzt'nte stabilit;,, is critical.Scinre DN.,\ llol\i.n.rases are inherentlv stable at 140'C- q,hereas

nranl' other erlz\'rllcs i.iie stabilized in r,rYo wjth unieue therrropro-ttct3nts. \4hen thcse iinzlirnes are separated fiont their proteatant,ihev krse their unique iherrrrtrstebilitl,.

Thesr cnzrnrrs rrra., have intportani applications in mernlne prl-cluctjon. nreiai leaching and rel-over,,. and in irnrnobilizeC enzyme sys-

lrnrs. in atidrlroir. the prissibi litr til'selectivc stereochentical mtxlifi cationrtl conr-Lrltntls noiliallr not in solutiort al Ltrt'cr tempcratures inay prr)

\ i(lL- n!'\\ r(ruic: firr directed chernical sYnthcses. This is lut excitrng alde rirrLnrlinl arell !11 th. nrrxlcnt biololicai science: to nhich environnttit-tel nricrobrol0gjsl: cul.r riluke .ignitlcaltt contnLrutions.

Estimated Total and Known Speciesfrom Different Microbial Grou

E sti m atesin V;:ious

of the Percent "Cultured" MicroorganismE nviron ments

EstirrratedI'otal Specirs

Knou n

S pecies''I)erce n !

Knorr n

[-nr ironnte^1 f-sl irnatcd I)crcent Cuitured(iroup

Viru ses

.Archaea

B rLc teri:r

Fun-ei

130000b

40 01Xlト

150003060000

5000<500480069000400()0

14]ビ

1121

う

67

Seawaier

Fresh q'ater

Mesotrophic lake

Llnpolluted esiuarine s,arers

Actir,ated sludgeSediments

Soil

()001-0100

02501-1001-30

_11_15 _′

02503‐

'Mid-1990 values and sh()uld be rncrcf,s€d li) 50!7.hThese values re sub\untiail\ uildereslimnred. p€rhaps b) l-l ordr.\ oi rnaqniludc

'l I indicales fiat these yalues ee probahl\'!!ro\s overeslimatei.dSmall subunl (SSU) rI{NA dak indicxre rnu.h hrsher in \nu di!enrr} dd Si.cn h\ cukure h:rrd sludi.\

Adapledfrom:D.A Cos3n 1000 \'ri.robi!l genonr\-theuntafrprdresourr fiSrr,cli Il:lJ-l6Table l, p. 15.

illcsc Prc、 1()tl、 l卜 ttllctJitllcti 11licIぃ bc、 11]、 ,iticiil〕 ゞthc Llヽ Ct)1)ヽ i11-

glc― ccH =cl 11liCrぃ c11ご 〔lI)｀ tll(ltiぃ Il を11)IP「 いtiChC｀ lhtil llllぃ 、、 :ltill iて ,111

(11■'tlsi()1l alltl nRicrt,bitll さぃ:]lll]t‖ liCatiぃ n 、、1lilc :1〕 illinli/iln g 、)、 'cl―

gro、、ith b) 11〕 o「C rtllDill―

=「

o、、ing c、 )11]I)Ctitい I、 1 ‐ ‐「 1 11‐

Genetic Manipulation of rh/1icrOOrganisms

Gcnctic lllaniplll(lti(Dn、 arc u、cd lo producc i〕 ]icF()Orgallis11ls、 1ヽ111

nc、v and dcsirablc char[lctcristics Thc clas、 ical nlcthods of llll―

cI()bial gcnctic、 (、 どで'(/?て り,′て,′ ノマI Pl(1)tl、 ittil rt)ic in tI〕 c dc、 ビIぃ P―

11lCl11 01'ctlittlrc、 lbr irldヒ lstritiI 11〕 icrobi、 ):ぃ=、

MutationO1lcc a prollli、 illビ ごultulc i、「 しlnd [1、 Liriピ l卜 ぃi tcCi〕 n itltiき ごヽti:う 卜C

uscd for culturc illlpro、 cnncnt illciLItiillご ご1lCrllictil l■ utagcn、 [lnd

Source: D A Coq in l{I\). \4 i.r ot,ial _cenonres-the ur lapped resource. Ibrec, I 8: I 4- I 6. Tsble 2,p. l5

tlltiti、 1()lci lighiぃてて■ノた1/Pr`′ ′ ノノI A、 a llじ 、arllplc.(1lc ril、 t culttllて

ぃ1 /'で ,″ ′(′ ///″ ″77■ ,「 tl子 |″ ,1 ヽヽh i t111(■ tll(l bc tlo、、11 01Rl、 tllldCrヽ tati

C01ltliti(、 11、 )iCl(lcci l●、、 tt,I]CC1ltl tlti()11、 ol PCnicillil]In 1913

、lrain o「 /t,′ ,ltア /ル ′`′

″ (カ ハ、て,マて'′ 7'′′7'ヽ aヽS iSOlatcd― straill NRRI

1951 -― 、、1lich 、、tl、 lullhcl in]PI o、 cd thlough lnutation(「Igur

12.1, ‐「 oda)ilno、 t pcllici1lill l、 lD1 0(luccd、 、.ith et′ ガ(1′ ′′′

“′7,(力 ′]ヽι

マて'′ i!`′ ′? gi■ )ヽ、n il](lcIぃ tDiCヽ tilrcd iモ 111lcntcr、 、、hich gi、 ics 55-1'ol

highcI PclliCillil〕 卜icld、 thtln thc origillal static culturcs

Protoplast Fusion

I)rotoplast ftlsioll i、 Il()ヽ ヽ ヽヽidcl〉 t.、 cd、 ith)casts and lnold`

1ヽ● 、1 01 lhc、 cl]liご iい い1=tl I]i｀ 111、 tll C.lヽ Cヽ tltll()r()f a sirlglc lllatinl

モ)1・せ ｀`hiCI(1ご こ:き ょ、きヽ 1:]ビ ごl‐ttilぜ こいi l ttinti()111 11ltlttltioll、 lh〔it coul(

lct`」 1(1、 ti t:ill tit=さ ilcl ti:い い Iて こヽa rl、 く,tit tcnctic、 tudic、 ヽヽith thcsl

1111ご [、 1● Iピ ti「 i、 111、 1■

1 1ヽ1 li〕 i tt、 1、 tii C PrCPtirCd b)i gro、 ving tllc ccll、 il

an ist)lonic、 oltlllon、 hilc trcatinゞ thcl11、 ‐ヽith cnzylllcs includinl

16 Chapter 42 lndustriai lr4icrobiology and Biotechnology

NRRL 1951[1201

1

NRRL1951 B 25[250]

|ド

X-1612[500]

|と・wis Q176〔 900]

|:

Bし3-D10

Pt:nit'iliiunt rrtqtte.fttrrii have been l'used r"'ith those of P' cltrt'so-

qettiun. Even veast proitlplast-r and ervthroc\'{es can Lre fused'

Insertion of Short DNA Sequences

Sho|t itngths ol chcnricallY si'tlihe siz':tl I)\'\ seqLrcnces catl be

inse rlcd intii re cipicnt qlicro(lrgenl.5111s hr tlle lltirce ss o1' stte-

directecl nrutagenesis. l'his can crciitc \lllllll gcilcllc ltltel lltign.''

lcaciinglitachiurgr\)1'Olle()iscrellt] atllitloltcirlsil)lhcIal('L'tplo

te in. Sirch ntin0r antilto liciC chrngcs itare br'rll lilLrtl'-1 itl lclld- in

lnan\ cuscs. to uile \Pcctcii ciletlgcs iil prolt'l;l chlrr:ritcristics' antj

itar.e t.esulietl in nc$ prodLlcts -sucit :i:; Irlilie cilr it'trnttlctllltllr re

sisl:iltt erlz) tllels atlti e llz\ lllr-\ thlit clin clitltir zc tle srred reactiot]s'-l-he'c apprortches ltrc ilal.t o1 tilt f ie 1tl o1' protein engineering'

49133P,23q

48-1655

49-482

49-2695

60-529

50-1247[1.506〕

48-786

|

i tl

48-1372[1,3431

1 ‐i‐

49-901

49-24291 Fi

50-724

50-1583

1 :'1,

51-825

1‐52-85

11■ |

52-817

11■53-174

:|‐ ||

53-844〔1,846]

llc,r.,are industrial microbiology and biotechnology sinrilar and

different?What approach is being used to increase the recovery of pre-

viously "uncultured" micrcorganisms from the environ'

i'n e nt7

What rs protopiast fusion and what iypes cf microorganisms

orr UsPi ir Inis orcr.::lDesci'ibe site-directeci mutagenesls and how i1 is us:d in

biotechnoiogy.

!!lat is protein engineerino?

3

)-2166

50-25

0-935

茸＼、

53-399 53-414

12,658] [2.580〕

馴fm ttf∞

馴翼

F

51-20A2 51-2033 F3121140]

F3‐ 64

12.4931

'igure 42.1 \Iutation \I:rkes It Possible to Increase:'ernrentali<)IrYieltls. A"gcncalogr'oi'thcttttltlLtirrlll]1 |LL'\\L'\

rsetl i0 inurcasc pc-nicilltn Iieltls $ith PdtitilliLtttl (l1t\'\()!(tltttlt

tsi.g X-t:uy,treilt.te.t (X). U\r trcat,iL-nt (Llyt. allcl llltlstard Sll\

N t. BY use 01,thesc lllulatioltal processes. the ficlci lr as itlcreas.'.1

l.ont 120 lnternarional L.lnits (lUtto 2.-580 iLl. a l0-1'old itlcrease .

-.innrarketl transfers u'ere usecl for nlutallt cro\\'th alld isolation.

fields in international units/mi in brackets'

_.ellullsc and heta galiictr,rrr,nidase. The p!otoplasts are therl rt'-

rtncr-atetl using osrnotic stabilizers sttch as sLlcrose Alter rcge n-

.:r-ririrrtt rtf the ccil ri all. rhe neu Prr:ttlplaslll 1'ttsitln prodttcl clltl tte

ir'eJ in furthcr studies.

A rIa.lrtr adr lntage ()1' the protoplast f'ustorl tecltlltclLrc i. thlt

protoplasts ot'dillcrent rlricrobial species cu be fused' er en il thcr

,ra nu, closelr lirlked tarontlllrcall.v-. For cxatrlple' protr-lpla:ts oi'

Transfer of Genetic lnformationbetween Different OrganismsNcr,, rrltcilltitrr, i.iare lrir,:n throLrgll tilc liilllslL'l ,rl nLlcle ii.lci.ls

l-..'1..'.rrtrtii1l'ercrtt (it !l1llt\ll1\. uliitll ir Irill'[ ()i tirt'rlipirllr 11t'r't'lrll'l

ing l ie1t1 Lrl ettntbinttorial hioloul t tablt -ll'-1 r l his inr trlr et tire

,r,r,.,.1a, r)t' !f It!-\ fl'ttttl tlll.' ot-latlisttt it-tto rllolhcl" gir rllg tile lt

i:iprr':tt rltitctl iellltitilitl.': .tr.li lt' ltll itltlce'cti ' l1t'ltitr t" 'lllrr,riti lrt.ltri.ltlilr,il de!tlt,l.ttl\rjl .\l llll|r)11"1 l '''ll\ c\iilllplc (11'tlli\

ap].,',rach \ a: thc rl ailil{llr (r1' th- 'stipcrbu!" paicnte tl b\

{ \1. f llikrrlribat-tr rtl l(lli. ri irich hrid itrl lllri'll\c(l caputrilih o1'

1,r,11,1.111irct1 .it!tltti;-itirltl. Th.'-lctle: lirt liiltihlotic plrltlLtcltoll call

lre lLl:l:1'L:l'letl irr:l itlliirrt'l-!lLilitllt l]llti lllLrtiriie::tilrrlllcrllliihi()ti!''r)1 C\tn ti) ii Iloil;11111f!ir111'-lllrl,lrieitl-r illiclt'L't-!'tltt'trl othct er

Itttlllie s ilrc tllc e\fi'\\i()n' tl !- ' "lt ()l ihL'

'll1) lllr !lt'11illrill't\'

tl-rrnr l)ir'rrrlr,rttt)tt(t.\ Pttiitltt lrird tltc ptiltlLtetirrtt o1 Pccliocill' a blc

tc'r-i0e tlt. ill a r cast ust'tl itl lr tl'tt' l'ertltelltltti0tl t" 't ihe Prtt lilre It

aor'tlr()il i n !' i.ltcteriel aolrl,tl I I I Illlllt \'D\.\ erpr.es:ioll tn tlitlcte-nI I rl--]llrll\lll\ Lilll llllPl()\t lltrlduelitltl

elllcicncr and ntinintize thc Pr-rnllu.rtir rtl rte P\ rcquiretl bctorc' the

prtltiuct is reat1l lbr r-rse , For cralllple. rectlttthtnant hltcultlr it.uses'(.\c(

lp. 10'1 5) can be repliclited in insect ia:-rae to achiere rapid

l:u s.' sctlc prrtiuclion of .L de :irctl r irui rrr pl ote i ;t. Trltnsuenic plants

r.1i,.71.r, i.,.,';'..'l', -i-':il1.r\hr'llt("11r)ll'.ilil'lr!liriilrl:j(qu'rtrl;-i 1.. ,i : ..:i .i\ ,.1 i;..:.:r..:. :'t ,.ii;.i. \lli"'' ll:l.L:;1.ll r' ''r :r' I !rl-

Llrll!ll irt '-. ';5i. f)\.\ :,li,.r l: Iri-lili i' irr 'lillllr 'll'rrrt 1l lil r'i\lil-l

D\ \ err:rtL'ri illiirt'ilt,'1:.l,la' ;iil!i il :lini'-!r'lil r\('i 'i ' 11'1i I t tl:'

.\,,rrtlr.;:tn!3 i)I ga!lal1! iill'trl'illelt,ln il15r) ai1ll ha ill5'l-i'Ll lntO

nlie fr)organisttls li:ill! \ iat(i1'\ lnd re conlbitlalll DN.{ tecllniqtir's.

47-638[9801 47-762

|

47-1564[1,357]

5120P,521]

|ド>＼、、、

52-318

' 嗜可~~¬

52-1087 1

425 Choosing MicroOrganisms fOr lndllstrialヽ 4icrobiolo9y and Bictechnolo9y

Property or Prodtict′I■ansferred

～licroorganism tised cOmbinatoriaI Pr∝ ess

Ethanol production Esc!rc rit ltia coli

Combinatorial Biologyin Other Organisrns to

1.3 Prooanedioi prociucrion L.. coti

Cephaltrsponn precursor Penicilliun tltnsoqenutnsv nthesi s

Lactic acid prrrCuitionXf iitoJ production

flrcatini n aseb

Perii oci n'

.{celone and butanolproti uct iitn

in Biotechnology: The Expression of Geneslmprove Processes and products

'\tttchurottt.tce.s t ereti-slae A muscle bovile lactate dehydrogenase gene (l-DH A).expressetJ in S. ceret,isiae..\. ceret'i:iae 957r xvlitol ccnversion from xylose u,iis obiained bt. irarsfr,rntins th; XyLI cene of plclla

rlrpilr enr'odin: a \r lose ,.J,u,,,." i",, a. , , "r,,;,,, "

,,,f ,,,, iUi. ,, .rrir,,, ,rl,],.i.,,,orgattistlt lbr the protiuction o1'rylito1. u,hich servcs as a sweetener in the tbod in6ustrr,.

E' crtli Expression of the creatininase gene fionr P.seutLtmotut.,tT;aiilri R:65. Cese inserted *,rth apt-rCl8 vechr. .,'

lntegratron tlf pyruvatc decarboxi'lase iind alcohol dchvtirosenl:c ll from Zyntomonus ttutbi! i.s.Introducrion of g:nes rrom the Klehsitllu pncutttortile ait re-gion into E. r.ctli rnltde pcssible

:rnaerobic 1.3-propanediol productiolt_Production 7-ADC rld 7-ADCA' precursors b1 incorp,ration rrf the expandase gene of

Ce phalosoyxt rin ac rentatiutrt itto penir i I I iutn by transfcx-ntarrun.

lirpression of bzrcteriocin t-ront PeLliotot.r;u.v ut itjilutriii in.!. rez,r,i.iiae to j;rhibit u,ineconlantinan ts.

Intrutjuctir'rt rrl; .hrrrrle \c\'r,,r r''1,, L.,tL.t I,t,tt',rr, /,,,. r, r,r rJrfr,,\(J (lrrrrl,lrilrrslufnr.,ironprotocoi. rvhrch resultecl i n ;crr,,nr- rnij bui.-r nLrl lL,r r l.ju!,n.

S. i ereyi.siue

C Iost ridium d( (tobutyI i( utn

一

，

守

「

ACA=?aminmephrlosfomnicacid:7,ADCA=Taminodecacetoxyccphalosporonicacid

t.t,.iue. lu:t ljio-l7 {5h \r1v'f i,JUr

Adaplcd from S ostcrsaa劇 :L01s、 Onland J Nieison 2(〕〕,MctabO:ic cnginccin8 of Sα εε′r7r77・ ●`′

∫ε“でドなた′ピ MicrObiol MOI Bi01 Rev 64(1た 34-50

\ e ctoi's l.ttt, scr'tirttr i J.-it jnclrrilr artif icial chr()ll()-\()ntL.s \ri.h asilrosc 1'or' \eltst\ (\,rCs). [rac.icr.ilL (BACsl. I)l bacleriophage,tit'i r c'd t lli'r)al.soire s (l),\cs ). a'd ,*1rlr,ilia, a.lif icial e hr,,nr()\( inlr\ (I,1.A(-r ). )'AC's ilre csl)ecilill\' \ arlLrahle because liir!cD\-.\ scqLren.c:; 1()\er l0(l ktr) (.a, Lre ,laint.inr.rl in the \.\f.ris I\eillltsle ait11rlnor0ntr tjt \ L,tiSt Ce ils. ,{ g0ttd erlr1lnle 0l 1gg1111.

Lr\. l\ Jlro\ idctl hr tire r lrus that causcs 1'oor,anr1 rlloulir r-liscascof iatilc anrl other liiestock Ge nctic jltlorrrlrticln tbr u lirot-and1l()Lrlli \1j\a t\c r irLrs untisCn calt hc incol.ir()trtled inlo l.,. r oli. l'ol-liirr ed t.r lirc r'rPreriron .i'this gcnctic infirrnmtic;n and sr.nthrsiso1'tlte .gent, Itr()(lLltl t iot i-rsc in vaccine Ilr()duction (figure .12.2 t.

(icnt-tie iitt.ritr;rlirrit tra,s1'cr all.*s thc. ltr.ducli,lt rr1. spe.i1r. llrotdin\ rinrl pr'ptide -s u ith.ut contJnrinarion bt siinilarirr.tlucr: thut nrrghr hc sr nrhesizttl in thc original or-sirnisrl). l-hisliPp,.rrrr1f1 aiilt dL.arr,.rsc lhc lintc lutd c(ist 01'rCc<trerinl and puriir rrg u Jrotlrrcl ..\nrtthcr ilta j()r t(l\ ailtitge ol'peJrlrde prodLlclionu rtir i-riotechnologr is tirrt oirl_i Lrioiogicallv acti\ c stcrc()is()rllcl-sr1r'c llr()duc.d. t his :Pcciticitv is rLrr-luired to avoid the possibleharrrlirl sirle eflecrs ()i'ir.cti\,e stcreoisonrers. as ()ccLlrrecl in thethal idontide clisasler.

What is combinatorial biology and what is the basic approachus€d in this technique? What types of major products havebeen created using combinatorial biology?Wh,v mioht one !.!/ant to insert a gene in a foreign cell and howis this done?\l/hy is tt i,nporiant to produce specific isomers of prcducts foruse in animal and human health?

Modification of Gene ExpressionIn ad,-litirirt trt inse rtrn! lc\\ ,jL-nc\ rn .rj:u1t.nts. it ltlso is ltLr:siblt to ntodilr gL,ltc iaqilllt jon br chalginr, ,,cne tr.in.ijl ll)tiirn. lu:ing llr()teins. r|r:litin!, hr br itl llr()nr()iers. lrnd re rnor in!1'ccrlbirck regullition r.()ntt ols. I hc\c liIlll()itr.llc\ ntekc it lto:.:ihic ltt or e rprodLtcc u rr ir.it r lir.iclr o1' 1trotlircr:. lis sho.,r n intable 42.-1.

l'he ,taclil'icatiiilt.1 gc,c r\Jlr-c\\i.n .rls. i;i. itc usetl [. i.icniiona]lralternrellltrolicpiith\\irrsb-t in.reirrationorder-eLulllronof'.[.1-!iljc !]unr-\. ',', hrulr is tlte f'ie ld o1' pathtar architecture. I-,n-clerstanding pathu'a! llrchitcLrlLrre rriuk:s il J)ossihlc 1O rlrsiln apalh\\'av that u'ill be ntitst ell'icic'nt hr rrrotciin_q sloriet-()r en.rg.ticallr niore costl\ routrs. -l'his arrpr.riiir hri: hecn urerl 1o ir.rpr.rcpenici J lin prorluction bv rretabolic pathu a-r €ngineering (\l pl._ r.

An illteresting cler elopntent in ntodil'r irtr Se rrr c\l)l-r\\lur.u,hich illustmtcs metabolic control rngincering. i, th;rt ot tlrcrittg c()ntrols lOt the svnthcsi\ ()1' l) copene . an jJIpr)riJitt iiltttr.,\ldant norrnnll)' present at hich ier cls in Iorratoe s anci tonrrrttrproducts. In this case. iin enqineerctl rr.gulalon circuit u a, clc

signed tit control lvcopene s\,nthe-\is in response to the inte rn.ilmetabolic state of E. coli. Another reccnt derclopntent is the u-se

ol modifled sene expression to produce varianrs oi rhe antibioricerythronrycin. Blocking specific biocherrical steps (figure 42.3)in pathu'avs for the sr nthesis ol- an :tntihiolic precursor reqr-rltedin rnoilil-ied final prrtrlucts. l-hcse rrite ; eti pr.rJLrct:. ri hi,:h hiir.-slichtl-r'ditlercnt strriclrlre\. ealt be ir,irJ irrr lheir p,r::ihlc :rt:-tinticrobial cftects. In ailditiirr. ir,i ihe Lr.c .i rhis appririi.h. ir i:p()ssible to better establish thc \rnrcture lltrctii.rn rrlatir-rnships ot'antibiotics.

VP#1protein

Foot-and-mouthdisease virus

Viral RNA

4

Viralproteins

Beversetranscription

Clone oi

ごVira DNA wlh

VP,1 9ene

謗¨I RettrctOn

l enZyme

r繹『鰤中

1

|

▼

不 :謂献

ll習∝mttOn d

Ａ

ａ‐ＲＮ円（ノ／ｔ、．ノ／

Ｖｉ

ｆ。

Foreign gene

Bacterialchror]]osorrte

ヽヽ＼＼ヽ、

VP11

protein

recomb nanttl acte ri a

Figure 42.2 Reeornllinant \accine Protit-tcti<ttl'

( r1' rer()tithi Itlltll D\.'\ 1c'cllrl tqLte:. I firrrl litltl t tlr itlih

aoaGa

30eVP#1 protein from recombinant bacteria{or use in vaccine Production

( itni. cotlinl tor ile:ircrl Jtroditcts can be e \Jrresserl in dilJ'ere ilt ()rsallisrlls. Iil' the use

.1i:,:rr.. \ 3.c jn. is prorluccd titroLrgit ciolting the vaccine gcne\ int() l:.tclterit-hia coli.

09鼈ラ

E^*rpl"r "f R".ombinant DNA systems Used to Modify Gene Expression

AcrinorhodinCellulaseRecombinant protein albumin

Heierologous protein

Enhanced gro\.. th i:il.".-\irlro aciri.:

St repirnnt t t s coeI itoIorClostridium genes in Bacrllus

S a c c h a ro t t t t c e s t- e re v i s i a e

Scccft arornr ce.i t' e ret i s i u e

,4.tpe rqi I ! u: nttlu litns(.',,t^,tt lu; t, t iiitti

Modification of gene lranscription

Arnplification of secretion through chromosomal DNA amplification

Fusiun tu a high-pruduction protein

Use of the inducible strong hybrid promoter UAS."1/CYCI

f)\'erproduction of glyceraideh-'-de-3-phosphate dehydrogenase

l:L)lxijL)r rri bio:r'nthetic Sene-\ rhat lead to enhanced enzlme :ctir ities or

r( , .Jr,'1 t:rJbr.k r.lLlJIl\\n

こゝ ostcttadrd:LO`sni mdJヽ た1%n2α"Mctab01で

cnsin∝ ing Or,ε「′口

"″)″∫εご″■5滋ξν:ε ,々夕ο′″ο′θiοι Ra■ 6“

'I)■50 Tablcl p 35

968

427 Choosing Microorganisms for lndustrial Microbioloqy and Biotechnology 969

ム

4ヽodule 2 Fvlodule 4

lヽodule l Modu e 3 lv4odule 5

Modified Structures

卜Иodule 2 4ヽodule 4 卜Иodule 6

Modu el 卜Иodule 3 4ヽodule 5X

3 ocked

einzy『 ln e

rv4odule 2 圧4odtile 4 卜4odule 6

Иヽodule l lt4odule 3 Avlodule 5

XBIockedenzyrne

，Ｓコ一】ｔ

Ｏ　ＯＳ聟ヽ「工一

Ｏ

C)thcl c、 tll1lPIC｀ illcltl(lc ti]c il]ビ rcti、 c(1、 ):]111c、 i卜 (,i tt1ltibiぃ tic、

al〕 d ccllulasc、 11,()(11lCtitiぃ 〕()1 ビc11ご じ` |)rCヽ i`t'I]l))ヽ へ al111っ lil'ictl―

11()13 grctltcr I)1()(cill )ヽ11[llC｀ 1｀ _ tin(1 11]1ご l tictiヽ C C11/)1llC ()ヽ Cllつ l｀ )(ltic

tioll o「 l c11〕 ●ヽ 111い 1 ltctibtit k i1lhilDit,ぃ :l Rcc● 1111｀ :Illlnl l)lょ 、1lli11ぃ ピc11

11)rc、 こ11111)lC 11｀ tl)(()111lD「 i｀ C ]()(o■ 1)`, ぃl thc 、、)l tl卜 Ic l)「 ぃicin ili ti

nlo(111lcd、 llaill a tc1litヽ 1(l iI]cl c(1ヽ C i11ビ ぃ1lCC1111を 11101〕 ()、 cl that ill lhc

oligint、 〕、tl‐ aill

Natural Genetic Engineering

Thc nc、vcst approtlch ilol crctlti1lg nc、 、 :1lCttib● lic capabilitics i11

a gi、′en ■licroorganisnl i、 tllc arc[l ol natural genetic engl‐

neering,、 vhich clnplo).、 frlrced e、 olution and adapti、 ie nlu‐

tations r、 どど/,″ 2∫ 9-■`', 1‐

his i、 lhc P!ぃ cc、、ol uヽ ing、 lDcc illic

cn、‐iI()innlcntal strc、 cヽヽ tい ｀「(ヽ rcc 1llicrぃ ぃ1■ tl i'iヽ nl、 1(, 111ヒ ItatC

tind をldtlPt thヒ 1、 cFCtitinピ 1111ご iて (`|「

=till i｀

子]]、 、ヽi〔 1l 1lc、、 hiい |ヽ iゞctli

CaPCthilitic、 1｀ hc 11lcピ Il till i、 In、 ●i tllビ ごヽ tidtiI)li｀ C nl tl〔 まiい ntll

P10CC｀ C`｀ incitldc Dヽ A「ぜtl lユ l11ご t i11● 1l t、 in、、hiピ ll irtin、1■

)｀ tilコ lC

clclllcnt、 and、 arious t卜 pc、 )`f rcct)1lRhirltllion Iつ i ll)critiCtil r()lc、

as、 ho、 n in tablc 42.5.

HO%′

一

.へ lthough thclc i、 llltlビ 11ご ()11[1て ヽヽCrヽ )CO1lCClllilng this arca,thc

lC｀ lD011｀ Cヽ いI 11]icl o()igtilli、 111、 1い 1ヽ1せ ヽヽ lD「

い`‐1(lc thc po(cntial()J

=CnCrtll l〕

g nlicrool=alli｀ 1ュ 〕、 、ヽith ncヽ 、 nlicr()bitil c1lpabilitics lk)「

tl、 c ill i1ltit卜 tl itl1 1]licI()IP i()1ぃ ぎ)tinti t)i()lcchl101()g).

Preservation of Microorganisms

(:)ncc ll nlicroo「 q(11l i、 11l ol 、l rtl、 1lt卜 bccn 、clcctcd or crc〔 itcd to

scr、 ca、 PcciliC purP()｀ C il intl、 t bc Prc、 cr、 cd in its original fornl

「or itll■ hcr usc and sttld、 Pcliodic tra11、 lcrs of cultllres havc bccn

t、 cd in thc ptl、 t、 altilougll thi、 can lcad(o lllutations and Phcno―

t),Pic changcs in lnlicroo「 gani、 1ln、 Tl)a、 oid thcsc Problcllls、 a va―

1lcI卜 ol culttlrc prc、 crヽ ation tcchnicltics lna)bC tiscd to lllaintain

tic、 ircdご じlttiTc chtirtictcli、 tic、 ltal〕 le 42.61 1´ }ophilization,or

r「cc∠ご_til卜 i11= tilkiヽ 1■ I tlビ ごiI' 11(ltliti llilliヽ

=cil ttiC trCqucntl).clll―

「lて いきti｀ i`tll 1llic:ぃ ●ritilliヽ i■ 、 へltht)ti=1l lぃ I`Dhiliztitiollを ln(l li(luid

ilitrて 、ごcl]｀ 1('I tlゞ C CtiC C● :い lDliCtit(:d tllnd rc_ltiil‐ CC、 Pc ll｀ 1｀ C Cquip―

nlcnt thc、 do tlHo、 、 11liC1 0bial ctilturc、 lo be storcd ft)r ycars

、ヽ1lh(ヽ Lll ll｀ ヽヽ ぃま｀、itl卜 ilit、 ()r til][lCCunlultitinil of nltltations

OO

O

lv・ odule 6

SS^- n:::--<".<O:.=

).,'rr HHo,.,. r> o, ---<HO> HO', /

', .<

" HO>i)

HO"

O

HO“

HOr,,

O

HO‖ "

HO HOJⅢ ‐

HO

一

DEB

970 Chapter 42 lndustrial Microbiology and Biotechnology 428

Natural Genetic Engineering Systems Bacteria

Genetic Engineering \{echanisms DNA Changes Mediated

Localized SOS mutagenesis

Adapted 1ia..nes hi iii n g

l-n-i.'1-n9. Tn.i 0 precise excistott

In vivo delction. inr ersion. lusion. anC duplicationftrrnration

Tll" II topoi.rn]cra5e l(cunihincll,,nSitc-specrl rc iecombinatittn (tvpc i trpoisomerases)

Trlrnrpo'ahle ricrtrents r1116r rpecir:r

Dl'NA uptake (transforrnation competence)

Base -substitutioirs. iranreshitis

i frameshil'tingRecipriral recon.ibinaiilr, oi'1lanking 8/9 bp rcpeais: ,e\iores orlgin,ll sequettc.

Cenera111, recip:ocll recombinlrlion of shorl sequei-.ce reireal\: occasionalll nonholttolrgou:,

Deletions anC fusions Lrv nonhomolotc)us iec()rllbinatjon. sometimes at shorl lepeu'rs

Inse(ions- ercisions/deletiitns. irr ersicrns bY concerterj or succcssir e cieaYage ligati(ln ieaciir)It\ Jt

\horl \(qllella. Ien. JI.: i,,icrll(. ;itt.;t.:tlchc.Insertigns, 11ansnositions. rcplicon 1lsi1ns. arijacert deietions/excisions. adiacent inr cisions br 1ig:iticin

of 3'Cll transp.r ton ends of 5' PO. grouirs llom silggered cuts at nonhonrulosou. tarscl .iLc.

Uptake o1'single stranrl indepcndrni ol se(lllcn.c. or of ri<;uble-slranded DNA carrvin.t spc, ie.identillei'sequence

Adap{ed from L A. Shapiro- 1 999. Naiu.al generic engin€ring. adaplive mutalion. and baclerial .volution. In MiL robial EL tios\ oI lnfedit)us D isedt?

for Microbjology. Derived from Table 2, pp.263-64.

E Rosenb,:rg. editor.159-75 washinglofl- D C: Americrn Sditlr

Methods Useci to Preserve Cultures of lnterest for lndustrial Microbiology and Biotechnology

■lletilod L'0mmerlts

Periodic transfer

N{ineral oiL slant

N,linimai metlirrm. clisiiIlerl qater. or'.\nter agar

Freezing in gros'th nredia

Dry uig

Frrrz--Jrr rrrl,.ropl:rirz;rl.,r, t

Ultrafieezing

Variables clperiodic transfer to new media include transfer liequency. medium used. and hoidingtemperature; this can lead to i ncreased mutation rales and production of i'ariants

A stock culture is grown on a slant and covered rvith ,rierilized mineral oil. the slant can be stored at

refri.gerator temperature

Washed cultures are stored under refrigeration: these cultures can be viable fbr 3 to 5 months or longer

Not reliable: can result in damage to microbial structures: *,ith some microorganisms. however. this can

be a ureful meanr ol cullure lnrinlenance

Cultures are dried on sterile soil (soil stocks). on sterjie filter paper disks. or in gelatin drops: these can be

stored in a desiccator a! retiigeration telrperature. or frozen to improve viability

Water is removed by sublimation. in the presence of a cryoprotectiVe agent: sealing in an arrpi:le can lead

to long-term vrability. u'ith 30 vears having been reported

Liquid nitrogen at I 96'C is used. and cultures of fastidious mrcroorganisms have been preserved ibrmore than 15 years

4.

1. What types of recombinant DNA technrques are being used to

modify gene expression in microorganisms?

Define metabolic control engineering, metabolic pathw/ay en-

gineering, forced evolution, and adaptive mutations.

Why might natural genetic engineering be useful in modern

mrcrobial biotechnology?

What approaches can be used for the preservation of microor-

ganisms?

MICR00RGANISM GROulTH INCONTROLLED ENV:RONMENTS

For nrani industrial pr'r)cesscs. inicroorgani\nrs rrru:t ht gr,.,"i n

using specificallr designed rredia undcr careftillr contlollcilcondiiions. including te mpelxturc. rerlti()n. and nutrient ieed-in-e. The developnrent of appropriate culture nteCia and tlle

!f()\,, 1h 1)1 rliar()()r:-iini.llr\ Lmder rrlrltrrtriril etlntiltiotls llrd the

rLrbicct' r)1' lil\ \aaii!rnI3.'iolr'1,'1 ,,.'1'q',lrnt. rl i\ naaai\:rr\ to eilrjtr lailltilloi(lg\. Thi

ict-nt ltrntttrtation. i:'i'ri tn lr Ph-r:iolLrgical \cn\L' ln arrrllcr-ra!ti\rrtl,rl i]lq l)1rirk. r\ anrIi(r\t'tl iir lt nruiir nrot't gettenrl ,,rar ttt rel:ttiott irt

intlLr.trrrrl nri.ir,broloSi linil hi()teahnolrigr. As noiutl itt table -12.7.

the tarnr alilr h:irt sercral nrr-rrning\. rneluding thc nrass ctllttrre (linricrorrrguni:nr\ (r)r c\en plartt alrd anintal cell\|. Thc derclopnlc-tlt

of industrial ternte nlation: rL'qLlil c\ uPprr )prl]lc uulture llle dia alld

the large \cale screening of nricrooreanisrl'rs. Otien r enrs are neede d

to achieve optillrunr product vields. il{anv isolates are tested lbrtheii abilitr t() \\'nthc\ilr u lre\\ prodLlcl in the iicsirecl qr,lantrt\'. Fcu

are succe<.iirl

Medium Development

Tite itie.liuill .iiiJi,.r--ji,i,\ -r iill!T(,,rr-g.irli.tll i.,IrLrr:il ilc-cllil:.- ltCltll

influence thc :iolrLrrrre crnrpetitir enes5 oi a p&liculi,u proaa\s. Fre'qucnil\. lo,,,,ct-a..1 anrLlr nlntcrillls are uscd i\ sOLlrces 0i carbon.

一蓼

Fermentation:

「

_

Microorganism Growth in Controlled Environments 9■

429

3

4

6

A hrVord with Manythe Microbio t

An-r prtice ss involvrng the mass culture of microorganisnls either

aerobic or anaerohic

1n,1' blological process that occuis

Food spoilage

source Ra,Materl11 _____―一 ― ― 一 一 一

~~~一― 一 ―

Ca:-bon and energv \'{olassesWhey

GrainsAcr icultu[a] wasies (corncobs) '

Corn-sieeP liqutlr

So),bean meal

Stick liqgor (siaughterhouse producrs)

Arntnonia and atln'qniurt salts

l- itratesDistilier's soiubles

Crude preparatitlns oi plant and aninlal protiucts

Crude ino:qi11llc chcnicals

Chirlk or crurie cal blrtlale;

Fcrtii r zer-graile PhosPhates

Hightr alcohols

SiiiconesNaturill e \tal s

LlLrcl artd r tgetahie oils

ヽitrogen

irr the abscilcc of O.

fhc production of alcchollc bcvel ages

Use of an organic subsuare as the electrr:n doror and a'cePtoi

L:e of an organic -\ubstl ete as a reductanl and of the saine partiaill

degaded orgallic substrate as an oriilalt

Gr,r.tth clependent on substfate-level phosphorylation

,1い ｀`ll｀ t()P10(ltlCtio11 01に lCSil‐Cd lllctabOlitcヽ

Grovヽ√th Of MiCroorganlsttSin arl lndustrial Setting

l'he O:.()ileelltliliioll lrn(1 1'lLl\ rillt llltl\! be 'lll'li'iclltll illgh tir

hl, e (l- ill .:rccss \\ ilhill lhe cells so li'rrit ii i: not 1;nritirlg. Tili: ls

arsllccilill\ ltl.tc $ ht'll ll de llse iuic|ilhjlrl 'illturd 't gl'1111 111g' \\'hctt

l'ilarncntttrrs i'r-ingi and iictillolll\ cctc\ lila itlltLllcille ration cilrl ilt-

c\r-n lul'thcr llrliitcd bl l'illrrllentoLls sr\r\\th ttigure'12'-ll' Such

filarle ntotts gtou th fe stlils itl a viscr'ti: 111rL:tie rlle r'liLlill' kno\\ 11

as 11 non-Ne$ttlnian broth' \\ilich o1l'ei\ d\cll IrrllL'IL-\i\i'1ll!( ir)

sti rrittg llll(l ilerllli()ll.It is esscntial t() lisstlre that Lht':e pilr 'lclil 1'lr''1or' lrlt- llot litll

i,i,," ,11!('rohiltl sron th. 'f his i\ tlt"'l ''riticril tirrllrll scaleup'

there a sLlcce ssfill procedLlr- de reloptcl rtl li strr'ill shakc lllrsk is

nrodificr-l lor use in a iaiqe 1'erttletltet ()t.ie lltLtsi [lnder\tiil]d the

microell\'ilolltlletrt tl1'thc'cultLire lttld tllliitltaiti 'intilet ctlllilitiotl'

neartheindividualcclldi:spitcitlcr"lise:inth':tltlturcrirlttt-ttt'I1'a sttcccssful irailsititln cllll b' tllade it()lli 11 fr()'L-\s ()riglnitll\

Figure 42'4 Filanrentous (irosth During

Fe-rmentation. Filatl'itltlttLr' ltrngr rrntl sctinolll\ celc\

can change thsir glouih tornl tlttring tile course ola ier-

rnentalio;. The clcrelopr.nent ol pelleted grouth trr

fungi has nlajor L'tl'ects on ()\\'gen transfcr ancl energ\

,"qii,",l to a!itate the culture' (a t lnitial culturc (b) \l-

ter l8 houls sro\\ th'

Vitanlin.,c'Ir()n. trace siilts

B uiiers

Antifbani ltgents

fvtrj* Co*Ponents of GrowthUsed in lndustrial Ptglgsses

けChapter 42 lndustrial Microbiology and Biotechnology

Figure 42.5 lndustrial Stirred Fe rnienlers. ra ) Liu::r 1ci'

illarllal' u.e,l b) ii ])ll.rr'nlrLtIir.a] it,ntl.:l|ti 1irItltt nticr0l.rl] itrr,-ducliOrr Ul',rnliltioirt.. rltr I)eitriir rr1 il 1L-rntL.nlat Ltnit.'i-lri. Lrnit

cen l.c nln urirlci lL'r'!)hia ,)L ;ilt.tL'r'()LlL conclitiort:. rtnd itutiidnlittltltit0tl:.:arrrltlrrg..rllrl tr-nIrlt1.t{ion nt0,,it0r-in! eltrt l.c caIr'rcdoul Ll|(la| 11\al)t rt e\rrldil j()ns. Il i()\rn\{)r-s antl inl l -tl crr nli rl| [{ rt-r ilreatt ttrttridc riitl linrt inlolntltlion ()lt thc c()uf\e rtl lhc ltrnterrlli-tiolr. Speril-i. :ubstrllts. rrrelelrolic iltlcrrnediate:. alttl linal pr-r,d-

LlCt\ rlill b.' rlr'tcclctl

tle rt'loptd in rr l-i{j ntl I--rlcnilcrcr lllisk tL, rr l0( 1.()0() litcr r::ii,tor. thr'n thc t-rrof C:r i)1 :alilaUJr hli. t.r,r,lt I iill-]erl ,rul f ritIe rlr.

\1re lrrLrulinisrn\ Li:it ire qto,.,, n r:t crrllurc lrrhcs. shakr. 1'llisks.

Itncl :liilccl 1c't'ttttnlcr'' ()i otitaj illlr\\ LLl llLlta \\ \Lcnt\. Stirrctl lirrllL'iiL.l\ eiln [l]l!e iir 'rze llont.l LI J li1.'r: to liX).000 Iitr-rs rtr'

llu'rei. rlcpcndirrL on lr'rrductjr)11 trrlruiL'ntclrl\ {{igure -12.-it ,\tr1ri1;11 indu:trial \1nrdd i'arntcnliitl()n unit i. illLi:trutcil in f igLrrcil 5/,. \r,t,url., ilL:st 1]1. nterliiillt he:l.,rillu.-tl lirLtt tar'.i11()n. i)HJcl.iir.lnteni. .:ilI1t1i n.l. liltil ltr rtc,:.s rtl0nilirIttlg lltLt\i bc cet-r-i,:il

r jLlt LrnrlL'r ri-rolouslr clntrolletl eonrliltorii. \\ hert rrqLrired. ioaltL()ntrol lrgcnt.\ lIr.r\t he lrtided. r.\JleciJll\ rr ith lrigh Itroteirt ntr'dia.Cttrttptrters arc conlttonl\ u-(ed to ntonitor outltut( f ilrn probesthut deter ritine lliu r(rhiel bio1111155. lr-r e is ol critical ntetah()licprod,lcts. p.H. inpul antj crhaLrst gus contposition. and other pa,ranrelers. Environntenllil conditittns can Lrc changed or hcld constant over time. de penclin!. on the -goals f()r the particLtlar proces:r.

Frequ.-ntlv a e I il jatl e ()litp()ltent in ihc titctliunt. olicn the clir-boir slrrree. i< li.i,l..il a()niinu()Li\l\ iontinuous feed.-so thatiht liiiarLrrril:ilti:nt ii tll ::,,i i:-rr i. i\!tL'.\ .;-th.ii.tii lrr:illlrhle ltl anrli',r-rl iittlt l-hi. i. !:1iir!ri,:1 lr 1;11|, ,:[.i',] .,t'ir Iil.,r:r- iind Othl-rJilrhlrltldriilai. II a\1i.. Liriao\a i\ J,.r'a\anl .ti iht br-sinltirr:: ()f a

1iL'ilrcntrltron. it can be ratabLrliz.-il to r ie ld e ihlnol. rihich is lost

Culiure ornutrient aciditicn Dissolved oxygen prol

Coolingwaterout

Air in

lu: l rolrrtiic lrotlLrr't uitrl r-etiLrces thc f ilril lielil. Thii clln ()cc1

i'\ rn rrndei ltc'r'oirit c()nriition-\.Ilesiclc: llte traciitionui stirr-erl aer't)hic ()r unal-r()bic f'enrenle

othcl lrpprolrchcs cirn Lr.' uscrl to gr(i\\ rrricli,rrganisnts. These a

ternlti.,cs. illristrrrtetl rn figure 42.6. includc irtt-tube i.nlenlertligirlr' 11.(rrr). uliiih clintinrite thc neecl li,r:tirrcrs that can L:

loLtled h,r'f illutcntoLts 1Ingi. Also arailahlc. js:rrlrr-1-state ferrnertrrtjon Iirgulc -+1.(r/r. irr * hich li plirticulatc \ubstrate rs keltt ntoirto nlrrntliin;t tlrin sLrrl'ltce riater i'ilnt uhcrc nticrtibes ciln gr()1

rnrl orvgen is aiaillrblc. In rarious l1'pes oli'i\ed, (l'icure.+2.(rc

and t-luidrzed-bed reactors (figure 42.6d)- the micrtrurulnj'nts lrassirciated u.ith ine'r-t surfaces as bittfilrns (see pp. 6(1.?--l). an

nredlurrj f-lou s past the fixed or suspended particles.Dialysis culture units also can be used (figure 42.6c.t. Thes

units allor.r toric wliste metabolites or end products to ditfusau av fl'om the microbial culture and permit neu,substrates to cliffuse thrtru-rrh the nrenrbrane tou,ard the culture. Continuous cultLrrc tr-chniqLres using chemostltts (figure.l2.6l) can ntarkcdlinrpror e ce 11 ouiprits liltri rttc\ ()1'srrhitrate uSt br-Cl1u-sc ntict-r)\)r!iini.ln\ can he ntlintained in a continut)i,ls iogtrithnlie phase

1-{ou cr e r. continLi(lrri ntaintenancc ol an ot.gani\nl in en litir,gro\\th phase is undesirable in rnanv industrial processes.

42.

ｅ

一鏑‐ｉｎｅ

972

pH probe

二

・ 42.11

{a) Lift-tube lermenter

Density difference of

gas bubbles entrained

in medium results in

flurd circulation

Microorganism Growth in Controlled Environments 973

(b) Solid-state fermentation

Growth of culturewithout Presence of

added free ',ater

(c) Fixed-bed reactor

l,4icroorganisms on surlaces

of suPPod material:

flow can be uP or down

(d) FluiCized-bed reactor

lv'1!ciocrganisn-ls cl sur{aces

o{ particles susPenCecl

in liquid or Eas strean '-upwarC:iovr

FlovJin+:

―Ar n

FloヽV Out

F orv out

SuspendedsupPorl Padicles

FoW n 一

(e) DialYsis culture unit

Waste Products ilfluseaway from ihe culture'

Subslrate may ciifiuse

through membran€ to

the cuiture

(0 Continuous culture unit (Chemostat)

Medium in and excess

mediLrm tc v/aste \Yittr

vJasted celis

Figure 42.6 -{ltcrnate \lethods ftrr }lass

(.JIture. ln atlclitiorr to stirrctl fr:rnlcniers' othet

lrr(llt',r1. . .ttt irc tt'."1 i(r L rlllrllt' llli' i""t j'tlll'ltlt ill

urr-ltt:tnril ilr()cas\ci' lti Inliilr 'ases

tilLlse altel'nale

lrnlll olichc' \\ il1 ha\ c 1(r\\ er ()lleratlllg costs alld. car

1,,.,.,.t. .irc'illiztJ !l ()\\ th'()ilditi(rll\ tlcecletl i'tlt

Ir rrriLrcl .\ 1lt1-la\i\

1ビ lalion、 hip to lhCヽ 'ゝ1lthCsiヽ ()「 cCll 11latcrial、 alld l10r,1lal groヽ 'th

lヽoヽ t antibioticヽ anci thC lllゝ COtO｀ ins fall intO this catC80ry

t. Uo* is the cost of media reduced during industrial operations?

;;;r;t iire effect of changing balances in.nutrients such as

tr4edium rn-*€

\ Iicr-ohi;i1 Pltiduct\ tlltcn arc cla::ifit-c1 as prilllllr\ and :ec'

ondar\ metabr)lite s. As si.torvn in figure t12.7. prinrar] metabo-

lites consist of cotripouncls reirtcd to the s\nthesi-i of migrobial

celis in thc grou'lh phase Thel inciutle ainirlo.ilcids' nucieotides'

and f.rrnentation entl orc"it'cts stlch 3s ethlirlol rrnd organic acids'

In adLiition. industriailr us'1'Ll1 L-nz\ 1l)!'\' e ithcr rrsttlciatecl u ilh

1|1g 11i;r1rl1ial scil: trr i\t'c1l/)111r" \'1t'll '11'- \\ilthc\iz'i fr iltr-

cr(rttt !:ttli:nlr,1url ilS !rL'ri tfi'

Secondarl metab()lites u\Li;illi ii!''tirlitllJi' jLititl: thr i-t-

rir-rrl ot nutriL-ttt litl'titatlon t)J \\ lirie pttlrii'tcl 'tiii:lll''illtitltl that iLrl-

lo$s the acti\c sro\\th phrtc The:e ctrlllplrllnds hare no ditect

minerals′ 910ヽγth factors′ and the SOtl「 CeS Of carbon′ nitrOgen′

and Phosohorus2\.^VhatarenCn-NewtonianbrothsandWhyaretheseimporlant

in fermentatrons?

3'DiscussSCaleupandtheobjeciiveofthescaleupprocess'

「

chapter 42 1ndustrialヽ 4i(robiology and BiOteChnology

5. Besides the aerated, siirred fermenter, what other alternatives

areavaiiabieforthemasscultureofmicroorganismsinindus.trial processes? What is ihe principle by which a ciialysis culture

What parameters can be monitored in a modern' large-scale

industrial fermentation?

system iunctions?

MAJOR PRODUσ S OFINDUSTR:ALMiCR08:OLOGY

Time

pre 42.7 Prinrarr anrl St'c<tridarl \letahrllites' I)e-

r,1ii,g ,,n tiic llalricLrilrr-otrllllll\llr' ihc tlt:ii'ril llrorlltet tlllti bc

,ra.al .1r.,ring or a1'ter o""tth Pi 'iirlrr tllttltboltlts lllLl 1(rlllle(i

-ing thc ,,ati, e gro'n'th ph:l-'c' u ile tclts st'c(rll(l'lr) lile 1'tbLrlite s

forrttetl atier grou th is cirllllllctctl'

Iniiustrilrl nrlctc,L.iolcigr hlts Ptt'rr idt'd prodLlel's that haYe in pro-

1'oLrrtCli' cilirngcd iir-rr li" es :rnri iiie -tpans The y include industrial

lnd ,.rglictrlrLrrlil prrtduclt' 1'ood lrdditir es' iliedical products lor

lrun,,,l unci rinjllii 1-rtalth. rtnd i-'ioir-rels itahle'12'9i Particularly"

in rhe iasl 1e',i r erirs- llonillliibi()ric !olllp\)rtlld-\ rtsed in metiicine

ancl healtit hure tliltrlc tilrijLit c,,tlitihLltiorls ttr the itnprrlved u'ell-

beinc o1'eililllal lrll,l hilitlrln polluf iiitlns' Onlr l-tllijor products in

r-i-re h arite!(rl-\ u ill lle tli'' ti'<cti ilr-rC'

Antibiotics

\lanr lrttiltrrrlla .lla 1)t'rtltltr'ti hi tllltl(r{rl!Jllislll\' preclonli-

ii1i1-,i]r lri iiiiiil(rill\ltl.ls :ll tht geitLts !/r1'r711r17;-'tt'' lrld b)'fila-

,,,a,,,,,u.'fLItlgt i.ii'. tirhlr' -i-i 'l' irl "l

' ; ehlllter' thc irnthesis of

seretlll i)1 tlrt'Ill(l\1 lllll'1i)llrrlt ltrltibitttl-: rrill be tiiscusscd to il-

ndustrial I'roductsEthanol (froin gluco-se i

Ethanol r:from l:lctose )

Acetone and butanol2.3-butanediLrlEnzl'mes

rgricultural ProducLsGibberellin-s

'ood AdditiresAmino acids (t.g . 1r sine t

Or-canic acids (citric aci'ilNucleotides\4taminsPolysaccharides

vledical ProductsAntibiot'icsAlkaloidsSteroid rran.'rfonnations

lnsulin. hunran grou th htlnlone' som3tosi'iir

BiofuelsH1'drogenN'lethaneE:thanol

-Sur'aftrrltriltr at': t {'r?1 1Jl(?('

K Itttt c rontt r t s .fra gilis

Clo st rid t Ltrrt ttce I ctbutt I i ttrtrt

Ent e t ol.tatt e r, St rrai ia

.{,t1rcrgi11iir, Bttt.iIIus l\1tt{ ir' 7 tt'-lndIrn1Lt

Ciltbtn'![o.fujLkunt

C o n ne btttt t' ri tt n 3l ul arn i t t tttAspergiLlus nlgr:t

C or : ne bat'le ri tt ttt 3l ut tt rtt i c tttrt

,\ :hb: tt. E re nu nlrc r'i ttnt B I ttkt sl t' tt

Xtutllrcntotns

Peni t' ill ium. St rep lontt.<:e s' Bac i I lt t's

ClavicePs PuryureuRlrr:,,r,u' 1'111"'l'a' 17r

Est Jier ichia t't',Ii \titcJruronu "s "/1'l lsl(l''

.;-., ith'r:,:' tr\A 'e' l-n 1 '':i

Phuir:r nlhctii miir,'itr:slisl:',\1 e r I i tt nt t b ti c ! L t i u nt

Zt'momon as. T he rnn onae ro bacl e t

and oth:i-s1「ltCl■ CrOnヽ

:' Prirnary- metabolite

な）

formation

Growlh ,/

、、

ヽ ｀有

; .." Secondary- metabolite

formation

Major Products of lndustrial Microbiology4213

（ＴＯｒｘ

ｏヽ〓くの）Ｃ〓〓ＯＦ一ｏ０

ぃ０ア』ｏＦＦＥｏ

０“０」０ゝ〓００」ＯＯ

．でｏ〓ヽご）のりｏＦ一〇一ｍ

o ?o 4A 60 80 100 12n 140

Fermentation time (hours)

Figure 42.8 Penicillin Ferlrtentation Iltrolres Ilrtcisc(ltlntrol of Nutrients. 'l-hc s)'rltllcsis o1'Perricillin hrgills n lt"rl

rrittt,-le tt 1't'titlt ltlttttlilt-tia becoltler lirnit;ng' '\1-tcr ltltltt ot'tht- lltc

t6s.. rlr sipl 1r cutahplizcd ilislrer'hlrt iclc)llri: fictlt dcgllded. r1ir

cr)sLt (it rlrpidll Ltsctl tlltlllrlslicchliritle) i: liddctl :lloll-l rr lth 'i lorr

icr el ,.rl- nitritge n_ Th js srintLtlate s nL[.inrunt tt litlsl'trrtrllititllr r)lthr'

cai-ltLrlt \otlrccs ttl llclliciliirl. Illt scllit t'alt()l i\ Irl tst'lrlttl tIitl-

the crtlt\etttii)n fec()illlllrrlldctl br t lle'\SNl"iilat l5' 1 11li111[1'r. t"t'i

the aris shoitld be IlrLrltiplii:d br'0.10 to olrtliirl tilc trttr'ralLte '

lustl‐ titc thC Clitical role of incdiuinn forlllultlti()n and cnvil‐olllncn―

tal contl‐ ol in thc productio13 of thcsc inlpoltant colllpounds 蠣ヽ

tll),i ti、 1 ,t、 、lit t lt■ 1ヽビ: 「` |

Penicillin

恥ぶ[1鮮議I勲熱、11#:Ⅱf鷺

i研硝‖篤胤烹博 諜∬だ11ゴ賓I葛Iぶ

‖窯∬)点‖I鷲∬fttIIT∬‖鷲Fζ‖ボヽ性R満樵mtibiolic icヽ ctt inぬ c culm「c hcginよ iヽn劇で■、ctngurC 42.9,un―dcl ct,11こ liti(、 11ヽ いic● 111「011Cd 1litlo=Clユ li1l111[ttiol〕

Anlino Acids

へ1ュ 〕i1lo[lC iti｀ tヽlch ti、 1卜 、111ビ [IR(1ピ 1ヒ ltユ 11lic tlご iti ttせ ょヽCd in tllc lkヽ いに1

ilndtl、 tlヽ aヽ 1l tlt■ tl● I〕 tilヽ upplclllcnts in blctld Prて tヽltiCiS tlind tl、 1lti、 or―

cnhancin=ご 、)11lPい und、 、uch t卜 1110n()ヽ odiunlピ lutanlatc t｀ lSGI

Figure 42.9 Streptonrlcin Production br

5tr-tpt,tmrctsgri-serls. L)tIlci"''- "i jiii''i':' lelii: 1o

nriL\iillunl antihiotic I icids.

０●一●

，

〓

。

３

Ｅ

ＯせヽっＥ

】

Ｅ

５〓ｏＯか

，一つＥＯ

（，

こヽゅ〓し

ｏＯｏ●ョ一ぃ

いＯ

ＣＯ〓●Ｌ”‘ｏ●●●０

奮Ｆ一ヽい、）Ｃｏ■ｏし″Ｃ●ＯＣｏ●

●一０、Ｅ

Ｏ″αｏ」ヤ∽

975

― ―

Chapter 42 lndustrial Microbiology and Biotechnology

ｔｅ

　

　

　

　

　

　

　

　

ａｔ

ｔａ

ヽ

１

１

１

‐

―

ァ

‐ｔｒ

＼、、、、　　　　　　軒『一′

a)

le hv Coi-r'rebntterirtt'rt qlulamicum. \ 1a.jor cer.hon tlrtws arc notcd by bold arrows(a)Growth with usc ofthc gi)()、 卜late bypass to

shitjccl holtj r|.ro$s) 'l he dlisiterl line-s ind;cate rL'lcti()ns ti-litt lre bcrllg used to a lesset e\tenl.

42.14

ｓｅ　　　　　ｏｓ

∞―十

ｐｈＧ‐ｕ　　　／／／／／一

ａｔ

　

　

　

　

　

　

　

　

　

　

　

Ａ

ｐｈ

ｌ

ｌ

十

ｃ

T百 ose pho

|CO

/ > Acetyl-coAC

CO

封r

)γ

」:te lileAcetyl-CoA

CHOC00~

Glyoxyl21s,,

′Oxa osucc nate

r

●Kao― ″・島

91utarate ‐

INH+

T

G utalnate ti))

Oxaiosuccinale4.

,,-n",o @*& UU.

g NH.**w

Glutamate

＼ヽ

Fumarate

I\\lr

Succinate

\ \Suce 1nYl-CcA

I

lsocitrate lyase

cis-Aconitate

\I

I- - - lsocitrate

&6sF

.!Irn() :iai11 pt'orittctton is t1'picall-r' cal-ric(i ()tlt b\ lllcllrl\ { )1

latorr ntutatrts. u lricit itavc lt redLlct'd ahilitl to liitlit svn-

. rr1 liir r'ttLi pir)rlLlal .f

h.' ltrlrnlal illicrool-lliillisrl ltr oitls tl!e r

l.ti(rn ()1'l-ri,rlhe lrillrl inlr:rnreriiltes b,r'the carefirl lcgulllli(lllllulur rlcirih,rli-ittl l'roduclion Lrf giutarllic ae id and scr eral

rllrnr)i1ai1i\i\Il()\\ al1l IiaLl outLlsrllgllltltxntsof ('rlr-t'llrl1.;ttt-

t: <lirtLrnitLirur tlllt leck. ilt htirc onl\'ll linlited ahilitr tcr

'cs. ti're lL \ crule IIttct-ltlediiite u ketog]Llla|Jle (.\ce dpPettr) t(-r \Ll.r'ln\ l-(lo-.\ as sh,.rwt.i it.i figure ;12.10. A ctlntrolled

riotin lerel uird the eddition oftattY acid derivatives results

:reasctl menlbrane permeabilit) and excretion of high con

:itions oi'siutamic acid. The impaired bacteria use the gl1'-

iie path$a)' (see set:lion 10.6) to meet their needs for

tial trrochen'rical interrnediate s. especially during the grou'th

'..A1ter gronth hecotle s lirnited because olchan-ged nutrient

:hilitr. rin lllnosl cott.tPletc nlolar conversiot-t (or 8I .79

tl a()n\er\l(ri) oi i\oaiirait t(l !luianlate occLlrs.

\ \iila. ilir a\scntiiil :lllllll(r llCltl usr-d 'ra) \ullPlerllent alefeal-\

rrc.1d\. '\ :is oriSinall\ ilrodu.e d in 3 t\\ o-step nlicrobial

PrOCCSS Tll:、 ha、 bccn rcplaccd b)を 1、 i llぎ Ic― tヽCp it「 IRlcnttltion in

、、hich t1lc l)actcritllll(｀ て,′l、′7こ′わで

`(rピ′′ι″77 .、 ル

`た

′′/111′ ″だ い1● ckcd iln

thc s、 1lthc、 i、 ol:honlo、 crillc ttCCtlllltlltltc、 1)ヽ lI〕 c ()ヽ cl■1=llitCI

Ctll]bC prOdtlccd iin t1 3(la、:fcrlllCnttltioll

Organi〔 Acids

(Drganic acid pl‐ odtiction by 11licro● Igtini、 l〕 1、 1、 111lPい lttillt ill i11-

dtlstrial 1lnicrobi(Dlogy and iHustratc、 thC cllicct、 ぃli ti tiCC I:3Cttll lc、 ―

cis and balanccs oln organic acid、 )ntlncsi、 and c、 crctiぃ Ii Citlic

acctic,lactic、 fuinaric,alld gluconic aci(ls arc nl轟 or prOducts

(table 42.10)Until nlicrobial processcsヽ crC dC｀ cloPcd tllC ina―

jor sourcc ofcitric acid was citrus tuit fronl ltalゝ 'Toda).most cit

ric acid is produccd by microorganisnls:70 r iヽ ヒl、 cd in thc food

and bcveragc indust]、 201を in Pharnlacculical、 and thc balancc

in othcr i1ldustrial applications

Thc csscncc of citlic acid fcrnlcnttitioi〕 i11、 |,1ヽ じ、li:lliti:1ビ thc

a11lounts of tracc nlctals such[1ヽ 111■ 11=tlll C｀ C t11こ l ir● 1l tt,、 tい

「 1ヽ―

/'ど′メ′/′″

'″′gど ′groヽ 'th at a sPccilliC lっ olllt ill thc icr,1lc1ltation l‐ hc

ゴヽ

'( > Aceiyl-CoA

CO:ツЮacdae / 6tde

Major Pr-oducts of lndustrial Microbiology 9774215

fVtrpt Orgrnic Acids Produced by Microbial Processes

Fermentation Conditions

Acetic iicid

Citric r.rd

Fumaric acid

Giuconrc acid

Itaconie lciti

Ko-jlc acid

Lactic acid

ism Used

Ac e t ob(1( 1 e r s'ith elhanol solutions

,4 sper.qi,'ltr.r rt i-ger i r: l.toias-ses brtsed

nredium

Rhi:opLts ttigrir:ans in sugar based

mcdiuin

Aspe r';illus /i!'gdr in glucose-inintrai

salts mediunt

..\;ptrqillu: lt'rrctts in mol15se\ salts

me d;un-i

,4 s pt rg i I I u s .fl tt rtrs oir'-rre incarbohvdr:rte tntlrgrnic N mediillll

ilomolirmentati !e a,i.1r,(l.tllll t

,it'l brur: t'kii

tatiYe Uses

Yr'ide variet-v oi' iood Llses

Pharnaceulictlis. as a lirod itdditir t'

Resin man,:lacture. talli'ling.:'rnrl sizing

A carrier tbr calcium and siidiurn

Esters can be Prtll''rterizedto make plastlcs

The manufacture ol fungicidesand insecticides u'i.ien crltllplexedu ilh melels

.\s a . rn-ier tor calciunl and

as an acidifier

Single-step ttxidation. *ith l5%solutions produced: 95-99% yields

I{igh carbohl'dr:lie concentrations and

cuntroiied lin.iitation of tr:ice metals:

60 80% vields

Strongll aerobic tcrmentation:carbon-nitrogen ratio is criticll: zinc

shoujd be limited: 60% f ields

Uses a-eitation or stirred fer:llenters:

9-i7r yields

I-lighli aerobic medium. below pl{ 2'2

35')i. vieldslron must be carefully controlled to

arojd rerction tirh kojic acid atier

i ernrcntationPurif iti mediunl used rrl tliilitate

eltracl iL)ll

Irrr-dirinr rtitctl i: treltlr'd utth totl d\uh3ll!e l.'tiil\ i!) 'll'tllc

l()\"

;jn(1 L()ltll!)llarl roitcr:lltflllir)11S ()i'lL\illlilllle tllCtlil'. illl!l l\ aillli'ri()ut in iiet.(lbia .tit rcd ielnt..nlL.t \. clerlL-rltilr. ili gtl \'l!iil L()ll! rrll

Ll.rii.'r.,ilr : r'r' rr'r'1..1.i,,'1'1,r11f i"ti i'i: i:' \''riri

Ierlct tllL i]lhii-tti,rrr oi eitrii li.id lll)tiLlctlon 1), il.t)il lrb.rt'().1

irprll. I hc stlrLt\\ Lrl thts lcrllltlltiitioll tie Ilt'nli: otl tht ltgtllltti()n

rrnci lirncti0ning ()1 titc ulr c()l\tic pilih\\ l\ lltlti 1ll.- Itie ltl ilirr) lit'

ilai(l !\cle 1ir.r' \{'(lit)tt !i.+)..Al'tcl tlle lLciirL'gl()\\1ll l)hli\'' \\lltll

1113 :11ir:1t.rrtc ler tl i' lirril. ciLI'ate \\ nthlst' ilLti\ it\ iilclclt\L'\ lirlrl

tltr- lriti" i11L'. {)t l.i(i)llliil\c illlLl i:0cilretc tlchrdro!e lllLsc dcrr':lL:c.-[]11'

1-sittlts irr iitiic ilaiLl ilucLilllLlllititln arlrl c\'1r-lioi'l hr the

sllL s.t'l lt,'' I ' :-.llri'll'.In !,)rtti.r:ii ir,rtl. iilt' llirrdLttlltrtl ()1'-Jlll'i)llic ilci(l i11\ rrl" l: lt 'itr

glc tlllr:rtrhilti cilzr lllr'. glLlcot3 o\idilsc' ilrLrnti tn l t/ri r'(tliii'

niqt'i: .1. rilqri l: !l()\1.11 Ltniltr (llltilltlllll coll(li1i(rll\ itl r '"111-.iealt jt(lu(rr ntctlir.till (rrtlirth bectltlles lirllrtctl br tlitl-i'!elt lintl

ihe rt-stiilg aall\ illlll\lorlll tiic lclllaillinS glLteose t'r !lLtcrrtlii 'tciil

rrl lt iril-!lt-slal) lr'11a11(r11. (ilLrcrltlic eciLl i: irsctl ii\;l (rilrri'l l'01 clrl-

aillill rlllrl i1(ril iiil(1 t( il Li)lll|()llr-llt 1r1 (ict'lgell1s'

Specialty Compounds for Usein Medicine and Health

ln atldition t() the trlllk pl odLlct\ that hare ilcrn prritluced tircr

the lasr -l()ro '1() \'cars. such a\ antibiotics' arllino acid\' and o1'-

ganic acids. illicroofganisljls alre u:ied ior the orodr'tctioll cli'

nonantibiotic specialt)' conlpounds. These include scx hor-

nlrlnL's. linlittllllor sgents. ir,noLthores. antl spcciltl '(llllp()ulld\

ihlt lnllr.icnce bactciia. tunri. atlrrehee' in:!-ct) iind fllllts,table-t2.l1r lrr lrllLlt:c). ll i\ llL-!c'\t':l:' t' ir"dilil rrittl Iitrr\il'ihe r,t-,,ttLicl. itirdci iaicl'rilli .irn1ri,11ct1 cirildlli'rir' l1r:i\\r'lij illlll

'Lht:t;r-lr-rilia]lr iillPt'rie;ll a(lll'lll(rtiIlLjr fL'llth illc'-rrll'!; illJT ril 'l

rrihlt. r1ltr l'\ i r "l'lJlli'rll.

1 What critical Iimiting factors are used 1n the penicillin and

strePtorni'ti n fermentations?

What are reguiatory mut2nts and how were they used to in-

crease the production of glutarnic acidby Clrynebacteilurn?

What is the principal ltmitation ci'eated io stimulate citric acid

accumuiatron by Aspergillus nrger?

4 Give some important specialty compounds that are produced

by the use of microorganisms

Biopolyiners

Biopol\'mers ltt'e tllicroi-rilill-r' llrodr'rced p()1\ illeri' prilllaril\i

1.,,lrsli.:ch,iri,l.s. il\ccl 1ri gttrdii) the ilo',r chtrlctcristics oi iiil-irirlr utt.l t() scl\c Lls ccllill! llgcilts. Thc-se llfe eniplr'r\'ed tn ll-tatlv

.rrcls o1 thc phar tllltcctlticll ilild lootl jndLrstrie: .' I

.\1lelr\lli'ioiallpoivsacchltt-ldesarcuscdirsstlihilizers'forilrc .ii:pr'r:totl ()i lrilrliltllllte-s- lrs f ilrrl-iorllling agenl\' or 10 pro-

ir(rlc ',i lte t t e tetltiilll ill I at iou: products. Ptrlr''lccharicle s he11l

ntiiintuin thc te \turc ol nlitn\ 1'rozcn lixlds. stlch ils iCe Creitlu. that

1i1.1 511f-iect to tlrasiic telllperatLlre challges' These polr:saccha|rdes

musl rraintairl their properties urtcler the pH conditierns in the par-

ricLrlar lbod and be compatible \\'ith other polysaccharides' The)'

sl-rould not losc their phy'sical characteristics if heated'

Biopolvmers also include (l) dextrans' which are used as

blotrci .jrpalrders atltl ahsol'bents: (2) Entinia polysaccharides

rhat lr'. us.d in plinis: anci (l) polvesters' derived fronl

F)\(,tLlttni()nti.r rrlr'r,r'rr'au1.r, nhich are a feedstock for specialtl

p1;1s11;r. L'tllu1L,.t'tlrtci(r1-illril\. plodLlcdd bt'trt '1t'ctoboc'tcr

.illin. rr. it\..1 r\ ll 1'r,,id thicke rle r. Polvsaccharides such as scle-

i{r!lLtcllt ale trsed b] rhe oii intlirstrr as drilling nlud additivcs'

t,

97e Chapter 42 lndustrial Microbiology and Biotechnology 42.16

Nonantibiotic Specialty Compounds Produced by Microorganisms

Pol),ethers

AvermectinsStari ns

Enzyme tnhibitors

BioherhicidesI;nnr u nos uppressants

Anabolic asents

UterocontraclantsAntitunrdr agents

S I re pt o mr c e.s c i ru ru t tn n e tt.s i s

S. losalien.sis.\. albus

S. averntitili.sAspereillu: krreusPet, ; c1 1 1 1,r,, c it rin unt I

aclinomycete'

S. clavaligeru.sAc'linoplanes sp.

S. hrgroscopit.u.s

To Irp oc I adi unt i rtJ ! a t tun

.5. tsuknbaensi.;

S. ltt e ru.; t r4t i t' L t.s'

Gibberclla :roeClaviceps puqtureoS. pertt:et ic u t suhsp. rrir:.irrri,5. lteucetiL.r.:.5'. i rzr'.tpilo-trr.s

N{onen si n

Lasaioc id

Salinom,vcin

LoYastatin

PraVxstaii.

Clavulanic acid

Acerho\i:

B iaiaphos

Cy,closporin AFK-506Rapaml'cirZearaienone

ErgLrt alkaioitl:Dororubi c i n

Dru noruh i ei n

\llrrrJl\arll

flleornr cin

Coccidiostat. rumenal grorvth prornoterCoccidiostat. runinal grori,th pit)n.toter

Coccidiostat. runtinal grorvth prornoterHelrninths and ailhropodsChoicsterol -lorverin-g lgen t

Cholesterol iou,erine agent

Penicillina.se inhibitor,Intestinal giucosidase inhiLriior klccre:ses h,r peigly cernra and

irislvccride s]'nthcsi-(l

Organ tr:rnsplanrs

Orean transp)ant:Organ transpllrnts

Farnr anintal medicaii0rInduction of laborL anaar treeimcntC:ncer treatilenl( itnaei trcrtlntnlL ancr'r lr i'lttntcnt

Compound Tlpe Source Specific Product Process/Organism Affected

"Compac(in. produced by Peniciilium citrinurz, is ch.nged to prr,.asElin bv iln aclinomvcete biocon!crsion

Based on: A. L. Denain 2000. Microbiai biotechnology. Tibtech 18:26-11: A. L. Demaii 20{X) Phrnir.eltri:it!\ a.l!e scrA- L Dernain. I999. Seconciary metaboiism in bacleria: Antibiotic pathways relnlation and Iunction in Bi.,l,r(r t,! th; pni:at

,nd!^' meebojites 01 microor,ganirms_ .1/r/r. Milrr,rr(rrl..r. J \\'- Len-q.ler G Drcrs_ and Il a;. Schleeel.

I Butahntl. i):155 .{'61: c Lancinredibrs 617-51 Nr\r Yorl: Jhicnr

-l\antlltn i-r()lvntel\ eitl]liltce ()il feco\r.f\, br, trtrl.;, 1\ tnLt \\.ltL,rf'lr)odin-e and the displaceruent ol'oii. 'l'his usc of xaltthun sLul.lll'otlttced b)'Xurttltontotld\ (LuttlL\//t\. tcptc\L-itts ll iarsrt ]t(rtant ul nrarkct 1or litis ntrcr ob jal i)r()duc1.'l'hc cr cl()d.'\irin\ har,e lr uniqLlc \truulLliLt. it\ sho\\ n illfigure :12.1 I 1or cr c],clodextt ip. Ther' are cr clic gliSrr:rictri:r,ritlcs u ht)sc sLr!als iire .ioinr(i br o I .J linkagrs. C-'t clotlcrtiinrcrin bc use(l litl ti uirie \ariet\ ()f Jrul'lloses be.caLi:c thcrc err.ircll ntolccriles hind u'ith suhstances and tilodil'\, tlre ir pitr sicllIlr()pcrties. Iror crantple. cvclode\trilt\ rr jll incrcase tltc :olLrhil-itr ol'phlrrrrurce Ltticals. rlraluce the ir bittentc:is. unal nllisk ( ]tcllrlciil odors. CvclLtdcrtrins als() can ire usetl ir_s sL-lecti\ g 1lr1r1lrg111r

t() r-enro\c cltolcstelol lritrit eggs anri bultcr. t, r []tllcrt \]l IL.\ll orrr oridal ir)n. ()i' us stat ionart' pilliscs in gas chrontitto!i ltltlt,\

Biosu rf acta nts

Biosurlictants are especiallr,. itnportant lbr envir.ot'rr-r-rental allpli-cations where biodegradabiiity is a ma.jor requiremenr. Ther, areused tbr entulsitication, increasing detergencl,. \\,etting and phasedispersion, as well as fbr solubilization. These properties rre c.-pecially irnportant in biorernediation. oil spill dispersion. and .r,hanced oil recoven, (EOR). The ntost widelv used nticrobiallrproduced biosurthctants are gl\ colipids. Thesc cornl(runds h t\ cdistinct hr drophilic and hr.drophobic rcgions. and rirc c\..lldltidispr-rsine agenls.

-l-he-r, have bcen use d \\'ith the l rrr.,r: lrrlajr.-

and other oil spills.

cylpS\-o- /r-v \

s1 PA."q:l^{/) "' i t*Sia*o,bo+ \\'

K,on Ho-,l'o..\ ryo9,o2) )o+_ t/ "t -g o-" 5to) | {[r---,-r- oJ o F

o',AO.HJ

ir-Cyclodexlr r-

Figure 42.11 .{ f'r clodertrin. The \irlrcturc rrl ,,-r'., clodcrlrin.llrtrduct-d ltx I'ltt'prtt,tlrtt(k)j)uLr(,1. Thcrc als6 tue [] 1;,i -v iirrlts. 61'

sinrilar \tniclurc bLit Ialger: these cor.npounds ha\e iltan\ applica-tions in ntedicine ancl industrr,.

Bioconversion Processes

Pr i, rrtr.rir', rl \ ii )il \. :tlrtr k n, r,,., it li. nlicrobixl t ransti rrntations r,r bi<t-transfirrntations..rl-1.;Ilin,,r ih.rn-ac. 111 1111ri3;ulg...r,Jtr ii\ lhr- in-

-\iitron ili .t i:rdiir:.i i trr kr,to liiniti0n ()t tllf \iitLtr.iltl]t)ide\itturaliOn,11'a ci'rrtpler crclrc stnrcture. that are can-ied out br nongro,,ring

L979Microbial Growth in Complex Natural Environments

4217

?'.C:O

Ho- ^. I -l-,i I-l'\J_iMajor ProCuct

Figure 42-12 Biotransformation to \lodifl'a Steroid' H1

ilrox-v-larion of pr,lgeslcrone itl ihc llrr position b1' Rhr:r;7;lr'r rli-

gri,',,,,.r. -fhe str:r,rid i5 il15-'t-'lred in accttlne belor-e ldilititln 1o tlle

ftr.-r,,\\ Ir itlllf :il . ttltLtlr.

n-iicroot-garti:rlr\.'l-llL' ttlicttlt'lgal.iistns iilLls lict li: tlitlcatallsLs'

BiitCorrr e r:tons Itirr e ll-lltllr atlr .intli!e \ i)\ L-l ChellliCill prOCttittres'

A nltjor adr rLrtllige is sterctle ltr:tll jcai' the hirritrticljlr ltclir e tirrrtl

o1' I prodr.ict is rltlttlc. [:llu t'ttlcs lt]str cltrrr otll \ ci\ :i)ccillc rtiic-

tionr utrricr nttlt1 rrrtlritlirll\. jlltl l.it'l't t'.ilut tll'illtthlt tlltllCcttlCs

clrn ht- tftrn:1,1I'n.tcii. l- ni.t1]Lil:il hlrtene. llatill()lt]\aL'tL'\. \L'llsls.

;rntl ntoltl:her..hCtIir:tJin\.1ilr)Li\lr1|q,rl1 -i'l()ll\.'lheCtlzrtltesre spttnsrbJ.. l0i. thc:c !.(r11\ L,f\1()n\ .etl hc liitlltct'llrllllr 0l' t\tflice i

Iuiar. cells 1'1111 hL'1tlrrtli.iteuL jn Lr:ttuil iil !()lltillur,Lis cLtltui.l allrl illcil

dried lirr dire,'t ust-. rrl lllc\ .irrl bc pre pltrc'd ilt tttole sllceif ic uavs

1,, \.,rf\ ',rt1 ,lr'.tr,',1 r'.,'. ,',\s1'j"r''.\ 1)'pialll t iiliritl', 1-l:i(i11 is tht hrdrorr laitilll tr1' a stctorrl

rligure 12.D.l ln this trrtrrpll' thc s arct t"t6lgLrle ste roid is

dissolyctl lp aCL-t(r1c erlii ti'i.'ll ldded tg tl-ie rcactiOtt systell thal

cotttltjns thc llrcurtlrl n rlliclrlbil] cclls' -lhc cotlrse o1'the rrrodi-

iicetiort is tttirrtittlrtri. rlntl tl'tc f inll ;rrotitrcl is e rtraclcil lronl ihe

tllr'Lllllrll llrJ 1',tt tl tr'i.BiOtranqf'onltat ions Cllt rie 11 oLrl br i'lCr' L-ll1)'llie S (lr lntilct ll()n

g11ru tttt uclls tio hltr.' ljnlitatiotls' I{clie liotl' thltl occitr in tlre lrb-

rc,tc" ,,1 uirjrr' ittel:il.olisitl .'r ithtrLtt leclLicin!' llor\e l r)l :\TP

hcing ar arlablr- a()lltllltl3ll\ - ilrt pt itnal'il\r cxcl q()lllt tcitL tir lll\

(sr'(' .t('( 1lr)i r ,t -j i. l1 {l-P ol rdrlLlct:lllts rile letltlired' sn cnerg\

':ource \Llch 3 gl111 ose ]lltl\l ilt'sirpplied urlcleI carclirlll'eiln

tr olletl ttotttr()u th arrlltlili()ils

'l Discuss the maior uses for biopolymers and biosurJactants'

2 What are cyclodextrins anC why are they important additives?

3 What are bioconversions or biotransformations? Describe the

changes in molecules that result from these processes

MICROB:AL GROWrHIN COMPLEXNATURAL ENVIRONMENTS

\,licrr,hial ntzlnaltrntcnt ul\(l ci]lt be carried ()ut in colllple x natu-

tll cnr !;o111llr'llt: -Llail ii\ \\llt!-r(- st'ils' 0t high ''t':'rnic lll'rltt-r

conlillrlin! aLlillPrr\i\ u lltrt- tht phr:tcll 'rtttl ntllrliiilllui

conditiirn: i,r r1ticr,,bitrl -{i()\\lh (11n1()I be col1iPlLrteiv contrtilled.

aniJ a largclr Ltnknolrn tllt.ittbt.tl rrrll'lnlunit) i\ present. Eranl-

製ilcul」ねFli:だ11lliF[Ft札獣W‖llnl猟轟Ia鍵

附f凛 1薯 11風)lin」責1撃。∴篤H∬蹴∬t選尊plicationsヽ 11l bc discusscd in this sCCtiOn

31ode9radation Using Natural MiCrobialCorllmunities

IPIittF‖1棚新ざTTi朧瀞ぶ緊i諄寝黒:

prよ iou、 l、 (｀('('/'59イ ).nlincralization i、 lhc llanSfOHllation o「

()rgを llliC nlolcculc、 lo 11111lC'cti l()1■ ■ヽC()litillall、

itヽ tlS assu11lcd gi、 cn tinlc and thC a11110st inl｀initc

ェI:や|1鍵:1罵gギ寵:キ1llti11:|=ポ ilまま:1属li鷺:lT

Oい、cい ltiい n、 。f na■ 出 i mdヽ )nthctic orgtti… ■甲 ■nd accu―

1lRliatioll i1l ntlttll.llビ l)ヽ'il()nnlclltヽ .hOヽ cヽヽ cr 卜c=an lo ralヽ CЧヒIC｀ ~

ti()n、 ュ1,o tlt[l〕 c tibilit卜 ol｀ nlicroo「=ani｀

111｀ 10 dCgratlc(hescヽ tll ic(1

、ub、 ltHlccS tlll(l thC lolc()i thC c11ヽ il ollnlclnt{CItl卜 、 allaCrobic c● :l「

‖1墓1驚[‖ili鮮湛‖i鮒黒1灘‖淋l醐i鸞肺=思

(‖

織:I里‖謝ド(濯#寵l:起II‖ ll。 lliふ「llよ1よ

週lIIli椰‖IIt卜iぎ1織IIIII11螢 1戦[liヽ c(11「o lllヽ aヽtcrl othCl tヽtldic、 llndicatc th.lt h)drofcn cti11卜 c thc

OヽtllCe Of lctluctを ll]t i10r thc dChalolcllation of(liffclcl■ C111011-

1111ダ llII常 i′)|‖;1淵 11ッ‡||′ }‖lぶ オ ヽ`器

下|)力l

Hunnic iCid、 .br。、Inish polynncric lCSiducs t)f lignill dcconl

驀 輔 幕 鮒

md:鰹 °

鷲

JttlHc醍崚 mmm囃 ぃ rC mmJd測

dc gi atlati(), ol thc lllを li 11 、(1■ lCtlll C 01'nl(ln)pc、 tiCidC、 tlin、 l ollnC:

i鼻彙!1熙榊‖illttIIiI瀧iII珪

Chapter 42 lndustrial Microbiology and Biotechnology 4218

' 42.13 Biodegradation Has Several

ngs. Biodegradatit)lt is a iernl that can

I to cle scribe thre e rna.jor t1 pe s of changcs

rlecule. (a) A minor change in the func-

lroLrps altached to an organtc cot-t'tporrnd.

substitution of a h1'tiroxyI gloup filr a

e rroup. (b).Art :r, iulll brr:aking trf ihc or'

onriround inio orgattic fiagtllellts in sircil

hrrr rhe original rnoiecule could bc reciln-

.i. {c') l-he conrpletc clcgradation of all or

ompor.rnti to rninet lls-

(a) Minor change (dehalogenat;on)

Che ln ca structt,re

O~~l~||~C00H

24-D

G~~CH ~C00H|

α6……… …H6…(b) Fragmentaiicn

G6……… α6…嘲

(c) Mineralization

.ciGO…剛―…翻

Approxinrate time todegrade in soil

3 months

2-3 years

81ocked″ ,era pos tiOn

(b)

42 14 The l′ εrα Ellc・ ct and Biodegrtldation. 1ヽ11lo「

ll di■ cl cllCCS CtlI〕 hを 1、で illl10r CllCCl、 ぃ11 lhC bio(lcgl」 (1-

)1'chenlical、 Thc ″′てィ`′

Cticct iヽ ti:〕 inlPo「 ttl1lt Cヽ そl11lplc

」il)i dcgradablc 2、 1-dic‐ hlor()phcno、 )'acctic acid (2、 4-D,

cxposcd nlcta position oll thc ring dcgrtldc、 111 、cヽ 'Cr[J

(b)rCCalcitrallt 2.4.5-trichlorophcll()入 卜'aCCtic acid(24.5-

thc bloc‐ ked nlcta group can pcr、 1、 [forゝ c aI、

。PPO｀ Cd t。 11lc● 1lh・「

い、iti● n ti〕 ビc(ヽ :l]Pい tilld｀ 1`1l l‐ ごcic―

l ti nluch、 1()、、c「 rtllc ]hc″′`ra effect i、

、hlヽ 、、〔l i1l Agure

hi、 、tcl cぃ chClllical(11「 lcrcllcc i、 lhc rca、 on thatthc cぃ 11]―

nrl)rt lir\\ rt it,-'ilri.i.lr i.1 tll.llJrrl',r1lh.'ntltr ile i1l. iif ill :l.j-l)t.uitlt lt cltlrrr,.r. ir: thi,r11111r Ir-rrttirrn. trll hc llrrg-elr Llcrllrie.l inlL singlr \unrrar lil Llrrlrli:- 1. i.5 1t'ichltlrrlDhctlo\\ ileetic ai'itl.\\rthilri)n\tiill(rrllrrliiriIllr-tlllli)\i1i! l.rri11lltlrjstirltlt.'.lilslirr'\r,\ L-lirl \ cill \. llnrl litu> t: i-r:cd irtt ltrt,:-t.:ltt lrt rtsit e itttttol.

.\l inrpirr.illtlrl:Ir.l ititttlrttltgrttL lrlrl,-lcSterilillr.lrl ts tltc t-ecr.lg-

nitiort thlit IIltn', 01 ihi'etltlllloutlds that atc arltletl to t'llrirott-nreirl\ llrd chiral. (rl Ir()\\ess lls\ nttllL-1r\ lind Illttltledtless.

\ii.r',roILlittirltt\ ()ititt ilttl dcg|ltr-lc (ltll\ (llle i:otttct "i e ttti.l-\tltncC. tire iltll-r i\()ntcr' '," til tctlluirl ltl tlle ttlr itttttllletli. ,{i lca.t

l--it, r,l ht-tl,iii,1t: ltre.hillii. ThLr. rt i: ilillcal lrr ldtl li-re nel'bi-

e itlc irortrcl lhiit l\ c11..ti'. c ltttd lLi.i, de grltdliirl.'. [(uct':li t'.ttdtr's

irer c -ho',i n thut nricrohtal ctrltttitr-tt.titie s irl dillert'nt e nr iLon-

nre nt\ \\ ill cirtgirrcle rjii'te lcnt ellrttltiottlcrs. Chltll-re . itr .ttt irotr-

nrrnt:ll conciililnr airtl nutricltt:iipltiics c:itt rtltet tllc petterns o1

i irilltl l olnt rl.-.! tildlili(rlt\1i.r,;bi.rl .i)nlrllirtilics cl.tltltg.- ihr'ir chltlt teri\tl!\ lll IC

\ll()n\t ro tllt- lrrirlitirrrl,tf iilrlrgellie ',r trrglttlic -iLtllsll-lttcs. [1'l par-

liculel cLrnrptrirrrl. \uch li' l herhicttlc. is.rkler.l rr'Peaicdl\ io a

nriclohirrl crrrrnruttiil. lhe citt-tttttttrtit\ 3dili)ti and fastcr rates l)lilegre.letion i.rrn ()ccur a pr(lcLr\s oi licclinleiiorl (figure 42.15t.

The aiiaptir L' Irr().e\:i o11tn is so ci'1'ectir c tilrrt this cnrichrtlent

culturc-bes.'ti ripprouch. estahlished on tite Principlcs elucidlted

b1 Bcrje rinck \.st,t' p. l/ ) can be Lised to isolate organisnts r"ith a

dcsir.-d se't ol.crpabililies. For erantple. a microbial comntunity

can heconrc so cfficient at rapid herbicide deqraclation that herbi-

cidc cl1-ectii e rtess is dintinislted. To coullteract this 1-rri-'cess. her-

bicitlc. clin he changed i(r tirro\\ thc nricrobial ctttittttunitl.ollblllrn.:. ihrl. i'r i':ar\ ilt! ihc .'ttcitir t'llr'Si o1' tilt cht-rlliceis.

i)t'gi:il:tlt,,rt iri{iar'\.a\ ihat,rcCttt in rtlll. llso clitt L-.c uscd illl:rtSr' :-lric J:,ii.r,l.riiLrtl ir1'hrrirtlcltti,)ll\ ()! \I'3:ta\ ti(illl iigl'tcui-

tLllal opL-ratl(rris. in a tcrltirquc talleii land farnring. The uasle

Microbial Growth in Complex Natural Environrnents 9創4219

，Ｃ●ｏＬｏＯ）

ゆＣ一Ｃ一ＣＥ

Ｏ」０つ一０一０」０エ

Time

Figure 42.'l 5 Repeated Exposure and f)egradation Rate'

.Ariilition ot att herbii:ide to rl soil cln resull in changes ill the

dcgradatir e abiiitl' o1' the nlicrobial cotlltl.ittnitr'' Relative tlegra

ri:rti0n ra{cs lor an herbicide a1'1er initial lddition to a silil. antl af

l('t I('lrc:llc,I c\p{)\tllt lrr IItt rittttc .h'ctii'"rl'

Figure 42.16 lヽicrobial‐ l■ lediated 卜ICtJ Corroslon. Thc

lllicr`ヽ biol()tical con osi()n ofir()niヽ an]lo「 p10blCin Thc=raphiti―

zati()11()fi「 OI〕 undcl a nl、 l blcb()n tl]c IDipC Surll、 cc tl1lo、 1ヽ1liCr001-

gani、 ■、 i1lcluding Dど 、′``rr l〃 '′

ル,toこ ()ロス)dc tlnc inncr sulfacc

The nrethanogens. an inlpo(ant group of the archaea that can produce

metliane . har.e had ri'idcspread impact on the Earth. Dr'\pitc intensir e

research. neq' discoveries are still heing made concerning lhese nli-

croorganisms. Methanogens have nou' been found to cont'ihute to thc

anaerohic corrosir-rn ol st.rft iron. Previousll' the microbial group usu-

ally consiriered the rriajor ctllplit in the anaerobic corr)sion ploce\\

r,,as the renus Dr:-safbi iDrio, lvhich can use sulfate as an oxidant and

h1'cirogen produced in the corroslon process as a reductitnt'

Nlethanrtgerrs cail ilse elemental iron as an electron source in their

metabolism. It appear-s that corro'\ion mev occur even rvithout the

pre -qeircc o1 sulfate. ri hicil is requirerl for functioning of Desullitvib-

no. Rates of iron removal b1 the methanogens are around 79

mg/1.000 cm: ol sutlace area ln a 2':l-ht-'ur period' This rn',u nt't tttn'a.lugh rate. but in ir'lation tl ihe plannetl service life of metal struc-

Iures in muds atrLJ suhl,surl-ace :olls-po:sib11' l'ears and decades-

such corrosion can become a major problem- Continuous ellt)r1s to

improYe protectlon of trtrn structure-t $'ill he require'J in vie $' of the

r-lirer.itr ul tr.'n ., rr, dinr iili( rooT-:.ir:'lll'.

nrate rial is incor.lturated into the soil or allo\.'efl 16 ilirr,r' ltct-riss [he

soi1 sLrrface. u herc rit-gratllttiotl occurs.

It is intportant ttl enrphasizc il)at such degradation Pr(lcLrsses

tlo not alrr a),s reduce en!ironntentnl problerrrs. In I'act. tlle partial

degradation or modif ication of an organic compound rna)' not

lead to decreased toricitl. An exanlple of this process is the rni-

crobial nretabolism ol 1.1, I -trichloro-2.2-bls-(p-chlorophcnl'l)

ethane (DDT). a renobiotic or foreign (chemically s)'nthesiT.ed)

organic cort.tpound. Degradation relnoves a chlorine tunction to

gir e 1. I dichloro-2,2-bls(p-chlorophenyl)ethylene (DDE)' rvhich

is still of environntclttal concern. Another irnporlant exanrple is

the degradation of trichloroethl'lene (TCE). a u'idel1' used solvent'

tf thii is dc-gredcd urtdcr atllerohic conditions. thc dange rotts car

cinogen vinr I chloride can be * nthesized.

■Diodc,Ind(ltioll al、 、、 ご(11l lCti(1 1い 、、itic、 1)rCをld dalllagc、 a11(1 11-

nanciallぃ 、ヽcヽ lヽctttl c■ 11(｀ 1ヽい ll i、 tl l、 11liCtllal l)i1111)。 11を11lt Cヽ anlPIC

Thc l)licI()biallヽ nlcdi(ltctiぜ い1■ o、 it)I〕 ぃ1 1llCtttls i、 I)a]■lcula11卜 ごril―

ictll、 、hcrc iroll pipc、 1lrc tl、 cd il]ヽ tヽitcl l(,g=cd t11lacrobic C11、 iron―

nlc13t、 o: in sccondar)pctF(,lcu131 rccい cヽrヽ ProccSSCS CarriCci c)ut at

oldcr oil llcld、 IIn thcsc r)ldcr llcl(1ヽ 、、をitCr iヽ pu11lpcd doヽ nヽa、 c―

lics of、 こヽll、 to l]orce rc、 idヒ lを11 lDctrい lCtinl t()a CCntral c()llcction

point lf tllc、 'ヽatcr co13taiI]、 lo、 lcヽ cls of organic(1lattcr and sul―

fatc.anacrobic 11licrobial co:1lnnunitics can dc、.clop in l■ lst blcb、 ol

tubcrclcs(figure 42.16,.rc、 ulting in puncturcd iron pipe and loss

of critical ptinlPing PrCSSヒlrc lヽicroぃ rganis111、 that usc clclllcntal

iron as an clcctron donor tiu〔 i11=thc rcdtiction of COc in■ letinano―

gcne、 i、 ha、 c rcccintiヽ いccll di、さいヽ ci ctt (｀ Iicrobial Di、 ersit》

Ecolog}42.2)Bぜ じ(ltl、 せ●fthC、 lヽdき「こ:1=C｀ )「 intcractit)ns that oc―

ctlr bct、 ccn 11licIて 、oritini｀ l11、 tl■ ご l、 iciユ lS thc nccd 〔 dcヽ cl(,Iつ

$tratcgics to dcalヽ iヽth 11licr(ぅ いial c(ヽ 1l oヽ 101l PrOblClll、 1,clticalClr - CHC1 -+ CIIIiC - CH:

一 initial degralation pattern

MO●org甲 1甲 ,W“

'9ut

“CめⅢ籠Sms＼ :船董1蘇"側

Юb

出鍛ぼ＼■||,‐・‐■|'■ |||:

「

h4 = Monitoring wells

:|||

Figure 42.17 A Subsurface I'-ngineerecl Biurenrediution Srstem.iiirtl it: Jro\sil)lr nl()\cnrcnt. \utrie ntr and (l\\ ge r) (ii\ iiir. pero\1dd. ()r

!r'(rLlnci\\ !ter 1(r ])rOnlotr- nrLtre e liiciclrt i-lcgr':rdati0n r.rl'crtntlintinltnts.

涌ヽ /1｀/1｀

/1ヽ

=Contaminants

Itiirrritolin:, lrnd recoverr u,ells rire u\cd t(l nrtrnit,rr the plumeo1h.r ()\\ qL-n rclL'a,qing cornpounds) ar-c added to the soil and

Give alternative deiinitions for the term biodegradation.What is reductiv€ dehalogenation? Descri-re humic acids and

the role they can play in anaerobic degradation processes.

Why is the "mela effect" imporlant for understanding

b iodeg radati on ?

Discuss chirality and its irnportance for understanding degra

dation effects in the environment.What are some of the irrrporlant mrcrobial groups rnvolved in

iron corosion?

Changing Environmental Conditionsto Stimulate Biodegradation

Olten natural nircrohial conrnr,-uiitres *'ill not be abie to calrv outbiodegradaticn proces\e s at a desircrd nitr' due to iilniting pirlsicalor nutritional factors. For e rample. bioctegradation cilien uill be

linrited hr, loq ori,\.!.ll lercl.. Hrdrocarbons. nitrogc-r.r. phospho

rLtq. and othe r ne.'rie tl nLrinari\ :il.r, rnlrr he absent 0r ai arlablco1llr 111 rl11u. flLrr rti.r.. iirLi' rr:rrrtrnt iJia' {rl dr'rr:idllilt)it. In thc.e

alr\c\. rL ls nL'ac\siir v kr tlelclrnine the Iinriting 1ric1()r\. hlred or.t

l-irbiu''r lrnrl S jrelford's la* s. and tircn to sup|lr needrti nr:rtelials

or.nrririilr thc cnr ironnrcnt. i . . .. ,, ',i:, r ,i : . ,

\lr)-\t ()1 tlrc clirlr ctlorts to stinruiate thc dclra(irti,.e iictr\ i-tie: ol' rnie roor-guni:n.ts irrr olveil thc nrodiircation olr \\ aters and

soils br thc uilditron o1'()\\,gen ol nutrienls. calied bioremedia-tion rrnd slroun iri figure -{2.17. Coutact bctuccn tire nricrobcsantl thc subsllate: the pnrpel physical envilonnrcnt- nutr-icnts.

o\\'gen { in nrost clises): and the absencc o1'toxic conrpounds are

critical rn ihis rrranaged process.

OIlen it rs firund that the rddition of easilr melaholized or-genrc rnatter such as qlucose incrcases biodegradatrr-rn ol recalci-lr'ant compounils that are usuallv not used as carbon and ener-et'

sources bv nricroorganisms. This process. temed cometabolism,is frnding rvidespread applications in biodegradation manase-ment. Cornetabolisnr can be carried out by simply adding easilycatabolized orsanic matter such as glucose or cellulose and the

compourrcl to he degradcd to a corlplex nricrobral comrriunitr'.Plants also nral he used to pror,ide the organic nratter. C{lnrdtaholi:rr is irnfor-rlilt in rran\ dilterent birrde-eradation s\ \ie nt\-

‐■理

4

「

4221 Microbial Growth in Complex Natura! Environments 983

弊 C02

Process F unctiorl

Stimulating Hydrocarbon Degradationin Waters and SoilsEr;rericnccs u,ith rtil 'ttjll. rit itlllirla clli ilrttllttt'llls illlt'ttllttthcst principles. \\iltert uotkitlL rrllh tli'llcr:ctl hr,ltLrcrtlhoirt itl

the occan. contllct bet\\ cctt tht ttticrooiLltnisrll. th.' ltr clr,-tcltrl-ton

substrate . antl othcl e sse ntiJl ttLtllicltts illust bt' ttlltitllltitle .l. Ttr

achieve this. |3llg1s t()rlttlitrtll-! lrtllri.nt\ llrld iill 1r1.1)Jrhilic ihrdrocat-bttn stilLrhle') l)icpilrirti()rt lllt',t hecll Lrs.'ti -1-iri. lrthlilri'.1.Itas acceleretcci the tltslitlrtiltt,rl rliiltt-titt ciLti1.. r,tl .iie k. br .:()to'109i . in crttttpat isrrtt ti ith c()ltlt'rrl t,il :ll.k. u ilelr lllt lttltliltottltlnutrient,s \\'L're not lrvai lanic.

A nlljor challcnge ior thr' 1slll1rlirr1r r'..tr 11lr' l:t'"tt \rti,lr''ciil sprll. u,hich occurrctt ilt .'\llr:k:,r irl \llirch l()8(l Scr crltl drt'

tarcnl allploaclles \\er' Li\i'(l t() itt'tt'list bir"lr:!t;r'lrltion Illcrr'inclLtclecl nutt'je nt additions. chtrrticll d j\lt!-!'\ltttt\. hi()\Lll iil.tlltllaclditions. arrri the use of hicit Jrrr'\\tttL- \1..trll T]1c Ll\L'(i1 il rlli-crobiall)' prodrtccd Slr colif id crlrul:i1'i.'i i.trt. irrt'r trr helpiLll

Thesc bictretttetlieltott;t|1ll-i-rltihr'.:il'a ll\!r LlscLl itl 'Lrils lLllti

seditlents. Filreranlpl.'.:i Illlirltlc r\\r'|-\t.l:lc J\l ,rrtr\\ tltn ltt Lt'ed

to degrade PC'Bs in riret .cdittte nt:. Irirst. piiltiltl tlt-llltlogcnilti(rllof the PCIBs occur\ nalurallr urttler ltnrL-r(lt.riL trrnililioils Tilen

the rnuds are aerated 1(l proittolc tl.tt- cotlrplelc de gllttlaiion tlf thc

less chlorinated rcsidtie s ie haptc-r opening figurc r

Stimulating Degradation with PlantsPhytoremediation. or the use ol pl3115 to stinrulate the ertractitln.degradation. adsorptitrtt. stahilization or r.olatilizrttirrn lrl aL)lltllrll

inants is beconlinS an inlponant pill't (r1'hii'tleSr:lJ,rtj,'n rt','htroi-

og1 . .A plant proVidcs nulriL'llt\ th lt '1ll()\\ rrrlllt-i:ih" rlr'|l r\ "t.Lrrin the plant r(rot zonL' or ritizrrrpilere rfigure 11. I E ,. Pllr i,rlcrllcdiatinn lll-\g inClude:1..liint a!tltrilrLtIj\rr.t\ l(),ia"t,1.1.1::r,ri. i:-llitl,rhi

■■

■■||=

こ え

CI CIGOGCI CI

ing t. enhanCed dr:gt-adlrti0n rtf cOntnrrrrntrnt:. Thr tlrgftttllrtion rri'hcrltihlLlt tlbetlzctlt is shilull !: llll Cr:tlllp1t.

Phytoextraclion

Phytodegradation

Rhizofiltration

Phytostabiiization

Phytovolati Iizarion

Llsc 0l pollurlnt accuntul;tting planls to removenrelals or orgrnics iioir soil by concentratitrgthenr in the harvcstebl!r plant pans

I -- | 1'l ril'' :r: 'l r.. .. ii''CJ l' lc'o 'l:alll\m\ Ir'

dcLirrili,rrgliric prll uiants

Lrsc of pllrr i|r)t\ 1(r 3Lr\(lrb uld ad,.orh poilulilnts.

irrrinlr r,rrtnis- 1-r(ri| \i'iiler and equeous rvaste

stIeaJlt \L.\. 01 lllen1s irr reducc lhe bioavarlabilitl'ofpollulants

ln iire e:rr LaorilcrL

Usc ol planr: io r',riatilizc follutar.tts

Based on T. Macek: N'{. Mackova: and J Ktis.

in cnlironmental rcm&lialion. Br.rl.drkrl. .4/lOЮ ExPloitation of plants fc・ r the κmoval of ottanics

18:23-3■ P 25

lizをltion ancI 、:()ltttili/tlti()11 lDl()CCヽ Cヽヽ まヽ :n(ltCd in table 42.12.

Tl a11、 gc:lic PlをlI]ts I1ltl卜 |)C Cl]]ID()卜 Cd i1l Phゝ lorcnnediatioll Usillg

cloning tcchlliЧ tlc、 、ヽlth マ`′ャ,わこれ′ど′ll`′ ll(ヽ どで'ノ'ノ '「

ノゞ, イア9-Sθ、

/10(').thc″7(′ ′へを11ld′ 71て '′ B gcncs htl、ビトccn il〕 tegratcd into a plant

(1′`7/,た

ノ`,ノ

,、 ′` 7/7`ノ f`″ ,て

`)thtIS illをlking it po、 siblc to transforill cx―

tl‐cinCl)ito、 ic()rgallic lllcrcur卜 f()「 11lS tO Clcnncntai nnercucy,v/hich

is icss of an cn、 .ironnlcntal ha′ ard Rcccntly transgcnic tobacco

pla1lts ha、.c bccll cons(1‐uctcd that c、 Prcss tCtlanitrate reductasc,an

CnZ卜 ]1)e fl olll(1ll Cヽ Plい 1ヽ｀ ビーCicfra(Jing btictcrit111l thcrcby cnabling

thc tran、 ビclliti l‐ ]ttll t、 1、 dヽC」「t,tic nilltltc e、 1ご r and nitro aroinatic cx―

PI()｀ 1｀ C｀ ThcゞごInご t:C ti:l卜 i、1ぃ (ililCd IPItull、 rゞO｀ `in s()lutions of cヽ ―

Plい 1`｀

=｀thtttご ぃntI Ⅲl:ヽ itinl｀ Ctilinて tヽ llヽ 1ぜ Itiビ (Dthcr Plants ha、 c bccn

Phytorcmed iation

Chapter 42 lndustrial Microbiology and Biotechnology

Figure 42.'19 Copper I-eaching fronl Lou'-Grade

Ores. Tlte chenri-strr rrnd rnicrobiillosv oi'coppe r ort'

leaching involre ititcrestittl clrllPlelnentlil re:lctjons''l-he nricrobial contributioir is the oridatitln of lenous

iirn ( Fer' ) ttr flrric irln (Fer - ). LtptrtsTtirillrrnt.fe rrrt'tt'i -

,1rltr lutti relaled Inicrrltlrg.rtli:ttlt al't \L'r\ lctire in this

oridatirtn. The terric ion then rcacts cilelllicalll' to sol-

Lri-.ilize thc copper. The solublt coppr:r is reco"ercil t"

l chenrrcal reactiorl uith elcnlenlal inrn' lrhich rcsulis

r n :rn ('lrrIlclitirl .,,|1rq 1' fl rt ;Ii1111,'

r'nginr'elcrl in the s;Ltllc \\l\ to (lt-griille lrichltlr'rt-tht icttc rtl ''lt' i

i-r)n IIlL- rll ;1 I aol-lIll ll I i llllilt i li $ tlrl dlr t tle ctlt.ict'l tl'

Stimulation of fu'letal Bioleaching from MineralsIlittleachin\, ts tlt.. usc of ntic|o()r'gat'risms. u hii.:h pioilLicc rtcids

ironr reducecl sull'ut ctlltlllot-ttl'-ls. lil crcate acidit ctlr itulttlletlt'

that sttlubilizc desir.eil nretal-s lor i-eco\ cr-\" This apprrlaclr is tlst'tl

to reco\,er lnetals front or-cs ltncl nrininq tailings uith in.'tal lcrel:

loo 1ou, 10r ,cinr,lting. Bi0ir:rching carritcl otri bY natulal popLrla-

tiotrs rrf Lt'lttr,.sTtLrillturi-iike specic-s. Thiobtr'rllttr 1/lirrrr'izrlrtrtr'

encl rrlui..tl thiohaciili. l'orerattlple. ullo$,s recoierr of up to 70'i

r)i thc cr)ppci itl lotr gradt' orts. As siltlu n irl figure '12'19' this

urr olr es rhe biololicel ,11id'11ott ot'ctlppe r prL'sellt in lhe se ort-s

to producc solLrble copllcr srrlfate.

It rs apparent tltat italllre $iil assist in blorc-rllediation i1'riVen

a chance. The lolc tlf Illtrul-lli tttict-oorglttlisrlls lll biodcgradation

is nou hetter-apprccirietl. Arl excellent eralrple is thc receni

,,rtrrk.,,,ith the ren ret-slitile 1'ungus Pltattarrtt-Jtat'tt'tltnstt.slttt'

i.rrrir (\[icrotlial l)iversitr' & Ilcologl''{2'3 t

Ol.tenhiodcgr,.lllilrnlndlriodeterirlrationhalt-Ill.ti{)rlle.Jjtire citects. utid it becoiltes intportant til ctlntrol and lirrlit thcse

processr-s b1 e n!irotitlle ntal mallageurent Problenls include ull-

,n ant.,l degraclation of paper. 1et 1'uels. tertiles. and leather goods.

A global colrcern is microbial-based metal corrosiorl'

1. What is cometabolism and why is this important for degrada-

tion processes?

2. What factors must one consider when attempting to stimulate

the microbial degradation of a massive oil spill in a marine en-

vironment?

3. Describe the major types of phytoremediation What is the role

of microorganisms in each of these processes?

4222

Precipitationo{ copper

FeS04

4. How is bioleaching (arried 0ut and \'vhat mi(!-sbiai genera are

invo lved ?

5 What is unique about Fi:aerachaeie chrt'scspltrLtfil? What

does its name mean?

Addition of ttqicroorganisms to COmplexAllicrobial⊂ onlmunities

B(ヽ th i11 1(1い ぃrtit(ヽ 1ヽ alltl liel(1、 t ti(lic、 tittClllI)モ 1`lttヽ こ卜ecil llla(lc lo

、pcc(l t11)c、 i、 tinピ 11licIい bit)lい iゞCtil l)Ioccヽ Cヽヽ 卜〕こ_ldil,ピ kn()｀ 1`1(lC―

ti、 c llnicl otlrtalliヽ 111、 1(ヽ (ヽう ilヽ ｀`[ltclヽ ●l いllncl C()1111〕 :き `、卜ヽ tc:l'｀ [1ヽ

|ボ尋置I1111‖鷲‖》][||:1驀li三:椰1llcnt culttllcs For c、 tinlPIC COnlnlcrcial ctilttilc I)「 CPttratlo11、 arc

tl、 alltlblc to lhcilitatc、 llagc lolnlを 1lion tllnd io i1111,1い C`｀ Cptic tank

PCri'()「 11lallCC

Addition of Mi(roorganisms、″ithout Considering

Protective A/1icrohabitats

Vヽitll thc deヽ cloP1llclnt()1(hC Supcrtう tlゞ

~bゝ=｀ 1`(11lakrabait)in

1 971 tincrcヽ aヽS illitial c、 citc11lcnt dtic lo thc hopc that such tlil illl―

proヽ cd llnicroorganisnl lllight bc ttblc to dcgradc h)'drocarbon

囃 lTR:『 1∫ I:|:書 i::詰∬ 1111tl∫ lattLih∴彗∫‖cC‖

crobc had to sur、「iヽ c and functio1l Enginccrcd 11licroorganisl■ s

、、lcrc addcd toヽ oils andヽ atcrsヽ iヽth thc cヽ Pcctali(♪ n that ratcs Of

dcgradationヽ o■ ld bc、 linlし1latcd aヽ IhC、 cI]]iCIい Orsa1liSnls cstctb―

1lshcd tllcl11、 cI｀ Cヽ (]cncrtlll)such adllitit)nヽ lcd l■ hヽon-lcrnl ln―

crcascs in rate、 ()f thC tlcsircd `ご tiヽ 11) 卜ul i卜1ヽicilil artcr a tc、 、

da、's thc tllicr()bitll ct)11lnltinit)Ic、「t｀ 13｀ Cヽ ｀`きrtヽ ::ミ ,il〔 ir ill trcュ tcd

CuSoo + Feo--> Cuo+ FeSo.

2Fe2｀ S°J3+CuFeS2+2H20+302-'CuSO`+5FeS04+2H2SO`

＼

The basidiomycete Phunenrclutete Ll!.n'sosPori tr? (the scientitlc name

nreans "visible hair. golden spore") is a fungus rvith unusual de-grada

tive capabilities. TIrts organi-rnr is terltred a "i|hile ro1 fullgtrs" because

of it.s ability to degrade li-snin. a randonl-v iinked pltenvlprop'ane baseci

polynrerrc component of u ood (se c .sectittii 28 -l). Thc cellulosic por-

tion ofwood is attacked lo a lesserertent. iesulting iil the charactens

tic white color of the degraded wood. This orsanistn also degrade-s rtruiy amazing range of xenobiotic ctmpound. ,nonbiological foreign

chemicals) using both intraceilular and ertraceliular enzvnles.

As erampies. the iungu-s degradcs hetlzt-nc. toluene. ethviben

zene. and ry'lcnes (the so ciilleil B-l'EX conlporriltls). chlorinated

compounds such as 2.-1.5 trichlorocthl'lene (TCEt. rind trichlorophe-

nols. The latter are present as conl:tlllinants in lr orrd prescl i :ttil e: .intl

also are used as pes{icides. In addjtion. 'ltir':: chlrrl illllied henztilts

can be degraded u'ith oruilhilul 1()iLlene! beitlr plc-r.'nl F't'n tllt ill-secticide H-rrdratnethllnon is degratledl

Ho.'r does thrs ttticrLrtrrganisnt carrr ()tti \uch ttllt:i \1.plre:rtlr

rnosl dcqraaleliott oi'thcse xcnilhiotic alrllln(rurld( (\L.Lir' ,,! .r .ttll'' c

growth. during the seconciarl metaboiic lgnin degradation phase.

l)egradation of some contpounds inr olves important extracellular en-

zymc.s inciutling iignin peroxiti:ise. rnanganese-dependent peroxi-,lase. and glyoxai oxidase. A critical enzlme is pyranose oxidase.

ri hich releases H,O. lor r-rse br, '.hc ntan,uanese-deprndent peroxidase

enz\ rre. The H.O2 also is a piecursor of the highly reactive hydroxyl

radicel. which parlicipates in u'ood de-rradation.,A'ppa,ently the pyra-

nose oxitlase .nzyme is located in the interperiplasmic space of the

tlngal cell rvall. u.'heie it can functrott either as a part of the fungus

or be reicased and penetrate into the ri'ood -rubstrate- It appears thal

th!' nonspecrflc enzll.natic s,vstenl that reiease-' these oxidizing prod-

ucts degrades rnany' cyclic. tromatic. and chlt-rrinated compounds re-

lated to lrgnins.\\'e can e\pect to continue hearing ol manr nerl' acivances in

i.ork gith this organtsnt. Potentjali! valuabie applications being

studicd include grou,th in bioreactors rvheic' intracellular and extra-

icllullr enzlnres can be iraintained in the bioieactor lvhile liquid,.\ i1:r.s fl(\\\ pirst the irrmrtbilized fungi.

litltl cirlllrill \\ siallls. .\1lt'i ltlllllr tlll:il''l't'\'1Lil :1111'lllIL\ li \r' 'i\|,Lrnd thur thc ieck o1 cllectiretla:s ol 'L'lll ltldad ctllltllas riar

rlttr- 1O et least thtee lliat(rr\: ( I I tila llltlrtctirelre:' o1 l.rLrItltlirtr

!l0u n ntictrli)tlillllslll\ a: a foiltl \(rtlrae 1i)l |ladlltol': \Llill it: \\)il

Irrotoz(lir. ilt thc irlehilit\ ()1'tht-\t ltticic-d tllictilrlr!.inl\llt\ llr trill-tiicl tht .onrpouttLi\ to he dt-glitil.'d. lrlil il I the iailLrr.- rrl thc

ltrlilcd n-iicr(tol!attisrtts Ii) \Llr\ ]\ L- :illLl c(rlllllctt' q ith intli-gt-rlilu:

ntieroorglini:nrs. Such a rni-rrlii'icLi tllictoirtg;illlstll tlllir tr.'lts' f illo a()nrllr:1a lutd srirr i"t hccartsc trt'tht lttiilitionltl ilt.'ltt-tti hltr

,1rrr rcqLrirttl t() llllutttltilt thc crillr D\.'\.\tterlllltr hai e bcctl tllaclc ltr tllltkt .it.h l.lhr)l illt)1 \ !rr)\\'ll

.ultuie: rll()ra ailf:Lh]c o1:ulrir.ll ill ll lllttlllill cll\l1r)nlll!-llt h\

Snrrrjpg thcnr itt l(r\\ llutriellt tllttlili t.r sIltliiltL tht ]llitJilrrtullnisnr: be lorr' arldint thcltt ttr iirt L'tt\ il()lltlleilt 'l'ila\e tougllr'ilitlt"Irppr-rilcitts hltr r- itttl-.rpr tci Itlicrobiltl sltrr tr ltl ltlti lllllcli6tt slnle-that. bLtt hr1\c rlol s,,1.'.'ii lilc ilr(lhlL'lll. Ill Ir-ct-lli \clil\. tildrd hli\

i-rt'lr les: intcre\t rrt .itllplr lltldill-! llllrrool!lltti.ltts I(' r'll\ iltrll-nrcitts ,,\ ithout cort:rcitring tht rpt.'illc tlichr ol 1111q11rq'rlr it"llnrL'ntinuhichtlt.-r ltl etostttritc:tiltilirllctiotl. l-llislllislcdtothtlitltj ol natural attenuation. lr hich elllphltsizcs tllt- tl\L- l)l lllttLl-

r ;rl nriclt,bial conrtttutritie. in the tttvtrolltttctltll ll l'lll-I!clllcllt'i r1

poJ I utants.

Addition of Microorganisms ConsideringProtective M icrohabitats\'licroor-canism additions to ttatttral etlr,irotlrrents ciit) be nlore

:uccessful if the nticrorlt sallisnl is arlded together rl itI-t lt illicrtr

hribirat th.t lire> the i)rgani\lll phrsicll P1{rt'cti()ll':is *ell e:

irr:::ihlr \Lrlpl) il'jg ntllllL'lli\. Tili. ill-ik.'. ;i 1lr'::li-'ie ir'r lilr- illi-

-:,,,,1-tLrlr:nr i() \r1r'\i\e in s;ltlc o1 ill.'itltcn.t'a()lllpctitl\a Pic\-

'ul.\ th:,1. a\i\t in lirc lliiiural e ill irtlnttlelit. itlcludrng PrL-ssure

li\,rr llrotoroiln ltrr'(lator\ sllcit lls ciliaie.. lilgellates. alld ail)oc-

bec \lie rolrlLbitlLls ntar ht' eithci livillg rlr inert. i'r'''lrr' ''

I-ir ing Ilicrohabitats. Spccialize-t1 lir iirg nricrol'tlbitats inclr-rde

tlrc sLrrlace'o1 lt s.-crl. I ro(r1. or lt ltal. r',hicll. uith thcir highel nu-

ir icnr 1lu.r rete und thc chlittct iiri in jtial colollizlttion b1'the added

nricr,rorgltnistlts..:ltt I)r{)tcct tllc lldclc'd tllicl'otrt'tl'oln tht'fierc.:a()nrldtiti\ c condiliotls itt thc lllliural ctlr itt,lltlle tit. Eranlplcs ill-ilrrde tht- rtse rrf R/rt:r,lrittrrt liittl lJtir'illrt' iltrttirtgicn.si.t. In ol-der trr

rnsurc thii{ Rhi-t,!tirrnt is itl clo-qe asstlciatirln \\'itll the legtttllc.

scc-rli ltrc co:Lttcl u itlt tltc rtticrtlht Llsills an rlii-t'rrgltrlislll llli\ttll'c.r.tr lllti:rtltiturt is pilrcecl irt tr brttd ulltler tlte 'ccd $'here the ne u iv

tleveloping prinrarr rool rl ill ilalletrlltL'. Itt conlrast' llttcillLt.s

rltut irtt,itrt.sis (flTt i: piaced on the sull'ece ()f the plallt leal'. tlr thc

p1:int is.ncinecrc(i to cottteirt tile tll- gcncs that allou'the plodtrc-

tion of the loric pt'otein in sitl-1. once rt is in-sested. Alier ingestion

br tlic targcl orsanisrtt. tite toxic protern u'il1 be ri'ithin the diges-

ti\. tlrct rrhele it is tttost cllectile. ir ':" '' ', \':, tr i " :ii,,.i.!'iiLIl\ ii:ril I ;l

Inert Nlicrohabitats. Rccentl,t' it ha,s been tbund that nlicroor-ganisrns can be added to natural communities together with pro-

tective inert n'rrcrohabitatslAs an exanlple. if microbes are added

to a soil uilh rnicroporous glass. the sun'il'al o1'added microor-

Sanisnrs can be ntarkedlv erthanced. Other microbes har"e been

t,t.:err etl t() e rartc their ou n ttlicrohabitatsl N'licrrrtlrganislls in

42.24

What factors m;ght iimit the

addition to a soil or water, toability of microcrganisms, alter

be able to Persisr and carry out

Chapter 42 lndustrial Microbiology and Biotechnology

*,ater column orerlying PCB-contarninaied sand-clay' soils

rbeenobsen'edtocreatetheirou'n"clavhtrtches"bl'bindin-es to their outer surface s rvith extrpolysaccharides'

'l'l-resc il-

ations shorv that *ith the application o1'principles ol t-nicro-

eccll0gt, it nla1., be possiblc tO iI()re succcssl.u]il, I.llllnAge

robial cotlli-t'lunitie s in neture'

processes such as changes in bioiilm concentlation gradients. It

is possihle to it-]easure the concentration ol sttbstances from

nranv diffcrent environntents (table 42.13) Aoplications in-

clucle rhe detectiOn ol' ciuci)se. acetic acld. glutarnic acid.

ethaltol. and biochcntical orrsen d'-:tnand In addition. the ap-

plicarion ol biOsensttr.s t() rrer\Lll'e cephaloslr,rrtn. nicoi:rliu

lctil. lind scr,e i'lil B r iilr1liirs hlis bet'n de scrrbed. Ilccentl'v

biosel.;ors har e he en i131'elirpc.1 using inlilluilochenl:ca1-based

detectii)n s\st.ir\. l-htst ner'r biirsensors riill deteci pathog'ns'

herbicides. iorills. pr()tcillS. alrd DNA. \,1anr' of the se biosen.

sors lr. blised on the use o1'a streptavidin birrtln rL-eotnitir)n

svste1ll tTechniques & Applications 42''l)'

i?.epid edr tllaL-s lLre heing nta.lc ln al1 iLre as tlf biosen-qrlr tech-

nologr. The 'c lne 1Ll.1e llrir.lili lllrfril \ clllL.iil\ irl the siabilit)' rnd

.lrr"iril,n o1'lltcse units. r,, iiich are he illg nlaiie tllttre portable attd

sensitir e . \1i.rL,r:rgltlttsttl: allri rtlctllIroljles such ls glucrlsr: can

be ntcttsrtrc.l. ihLr: rllec-ting clititlal ngg{-s irl tllorlern medtcine'

M icrcarraYs

.\ lurt.' L]i11i ,ii tl'r. llt-\r :tllrl dcr ilitllirlg rrlie i obial biole lhn0lrrs-r

tl. l.' 't I)\ \ '(rirl(ll.(\ lll gt'n('llrril\\ lr'lllr'Iiliilt

!-llc t\|1f :'1|il ill r'()llll)]c\ biilltrllclll \\:i1'lll\ tr"'ir'r'llrrlt 1-! (;)

il,. ,"1..'.i iiLl\ il;rLL,. tlrer hltr d r.)r.cL1r1.r,ri lir thi, lire ir ai.L- tha re \uit

(r1 .r \ r!le \\ itt g.-:l,,tlliu.. i'atlol]lirilliilll I)\-\ tt'clln' r1l-'r ' optiur'

Itrtr,i;1.'\.-'rl,r'.irl.riill:i1']'' ''i:'t:'lr't1rrttii"11''I;i|;"'qt'

ing. fhis nticroarrav tecltnologv ila' hccll srtgucslctl to proVici:

thi cquir elL-nt (r1 lhc cilcrlli\t's pcriodic tlrbie lt ol1ers the potetl

trtl ol lis:ltr ills .ili gt-rlcs ttsttl ltl essellli-'le atl ilrganisnl anci cltn

nt,rttit0l crp|r:ssiiltl tll'tcns il1 tltrrLt:ands tl1'gt-'l.ies based tln lhc

pr.incrple:'l,,,.,,, i,, figtrre {2.1 1' ln tilir tt-chnitlrrc' 1 0() to l00 pl

,,,1t,,11e.. cLrtlt:iillillg tlt'sllt'ti \e(lur'il'L-\- llr' sP()l1ed tlnto glltts

-t]r.jt:rli!ltIld]j]lerttllltrct.iltlslrtitltlt.irlri.l.hes.:ltrtlitsarethetltllirlt-l.L l, it}l .|)\'\: 1]ll]11 JtIl( i,\ll|.t.\\](lll rr, r,il. .i/.i1- Birldirlg

o1' tlrt .l)\.\ lrrl \ llliirll\ -gctle: it tlrta'tircri trsine rlpitl photo-

l,r(lll! l;r,'lrit"l ll,: I(i lllllLll'(''

( irtttlltlciltl lllle r()iii-la\ ptLrdltc''t IIIL- ll()"\ ar ailable that con-

tair (t.iJJ iillcll lf.ltllt':1r)l 'LIUL'llllls gcllc c\Prt's-\itllt in '!rrr't'ira-

i ()ii 7\( r,\ ( (,r.(,\ /\i.r(,. F<tr [-- t'ttlr.1.l9(t6pg1 relldrlg llatlrcs ctn Lle

\alillllcLi itl li tltl.toiift.l\ 1Ul ill'11' l(lllt''cllilll'l 'he etltire 1-- r'o1r

desired iunctions?

What types of microhabitats can be used with microorEanisms

when they are added to a complex natural environmenll-

赫hy are`Iants inoculated With θaθア/′υf_1カ

」月βσ′θβSS?

t5 BIOTECHNOLOGICAL APPLICAT!ONS7:roorlianisttts and parls t't ttlicr"()l'Jlrni\lll\' c:pecialll cn-

lcs. ul-L. rrscil rp a ri ide r ariet\ 91'hjote chtl'rlo"1.xl illrpli.ilti()lr5

l1()nitorthe]erelsol.CfitiCalct,rllptlrtlit].itl1i.tc.:ilrl]()Illll!lll, in rrtirttal,s and hullllills. These tccilllitlLtc' hltr c rr ldc a1l1t'1iclr'

ts iJt el]\'iroillllentlrl scicncc. .inirtlai lltttl ltitlttllt.i ilr',tltll. .ltrJ it-'

ic science.

)5ensor5-apidll, tlevelopiilg llr.ll of hiiltechnolocl' 1i111gti t.ll i tltetrsr'

srnational scit'nlif ic irtterest. is that tl1'biosensor proriitctioll'

this lieltl o1' bioclectrorlics' livillg lllicr()organi\ttls l or the ;l

,\'1lres ol orgatleiics) are ltnked s iiil ciecilirtlcs' atld l-'i1rlo:-l

re actiotts ilre converted into clectlicel cttrrttll' bi tllr-'c

)scrrsors (figure 12-20) Biose nsors a|e ltcitlS ilcr c-lopttl t''

rasr.ire sllecilic coltlptlt'tctlts in bt'ct lI ltlIllil' rt POllLtialllt' 1Lr

icct f.iari.r- cr-.r.tp.u'cls in i'o9ii. a1d lo :tur-ir, r'r'rr irirlttllciltlrl

Molecule discriminatingfunction

誓逮癬（ＳｕｂｓｔａｎＣｅ¨ｍｅａｓｕｒｅｄ

E lec t ric

sig nal Biosensors: Potentialand Environmental A

Biomedical, lndustrial,ications

Clinical diagrrosis and biomedical monitoring

,A.gricultura)- honicultural. r'eterinalv analysis

Derection of poliution. and microbial coniamination ot uater

Fermentation :inalr sis and ctrnlrol

\'frniarrino ,rf inrJrr<rrirl oere< 'rnrl lirrrrirtt

RecePtor

gure 42.20 Biosenstlr Design' Bitlseils'rr' 11q- i'rr'l'tr.: rr'-

r:asine applicatiotts in Inedicine' inductrial 1111;1-irlrllrlrr'gr ' riilJ

Receptacle substances(enzyme, antibiotic.

Physical and

chemical change

〓一

J…Bi5tin Bindi and Biotech

Egt u'hite contains man-v proteins and glycoproteins u'ith unitlue prop-

ertie s. (Jne of the most rntcresting. w'hicn binds tcnaciou-sl)' to biotin. *as

isolated in 1 963. This glycoprotein. called avrdin due to its "ar id" brnd-

ing of biotin. *as suggested to pla-v- an tntportant role: making egg rlhitc

antimicrr)bial hy "ryirg up'' the iriotin neeilcd br'rlanv rr,i!r, u,rg'rttisitl:

Ar,idin. uhich fiinctions best under alkaline coniiitiot.ts. has the hishcst

kno',r'n hinding utrnrtl betueen a proteitt anii a lrgard. Scveral Iearslater. scientists at I'iclck & (-o.. Inc. discrrvet'ed n sirttilar protein pro

Cuced b1 an aclinonrlccit. SlrrTr,orini a.s uvirlirti. nhich binds bjotrn at a

neutral pH arrd u hrch dtv:s noi contain carbth)'dri1tes. Thcsc characler-

i,stic> nrakr: \treptavidin'rn ideal biniiing acenl li)rbi()tin- antl it has'r':en

T,arget : Binder

used in an almost unlimited range of applications. as shown in the Box

figure. The streptavidin protein is joined to a probe. W-hen a sample is in-

cubated with the biotinl,lated binder the binder anaches to any available

target molecules. The presence ard location of target molecules can be

determined bv treatinc the sample u'ith a streptavidin probe because the

streptavidin binds to the bitxin on the biotinylated binder. and the probe

is then visuaiized. This derection s)'stem is bein-e etnployed in a wide va-

rietv ol biotechnoic,rgical r:pplicatiorts. including use as a noffad;oactive

probe in hy'briCization studies anc as a cntical component in Diosensors

tbr a u ide range of environi,,cnl:rl ntonitoring and clinical applications.

Not had tbr a protern iionr a "sintple" filamentous bacteriun-r!

Probes

EnzymesRadiolabelsFluorescent agentsChemiluminescent agentsChromophoresHeavy metalsColloidal goldFerritinHemocvaninPhagesMacromcLecular carriersLiposomesSolid suppofts

ｌ

ι

ｒ

Ｊ

AntigensAntibodies

LectinsGlyccconjugates

EnzymesReobptors

nanspod proteinsHydrophobic siies

MembranesNucleic acids. genes

Phages. viruses, bacteria.s,,bcellular organelles. cells.

tissues. whoie organisms

Strept:r\ idin-lliotin BindingSlstems are Finding \1 idespreadApplications in Biotechnologr.\Iedicint., and En\ inrnntentalStudies. lliich IrolccLtie o1- strr-p

iar itlrn. r pr'()tcin de r ir ed il-ottt atr

rlclin()nt\ cctc. irl: lolrr site s brq hich it crn hirld tcnlciousl\ tr) hi-i)till lnolcrl jir lcrir. 13r.rtlaelrilg Ibinrlcr to thc biotin. and a proht-.

such as a f-lLlr)rescent nrolecule. to

tire streptar idin. the target rnolccLlle

can be detected at lo\\' concentra-

tions. Target brndcrs. probes. and

applications are il()ted.

AntibodresAntigensGlycoconjugaiesLecti nsSubstrates, cofactors, inhibitors, etcHormones. effectors, toxins, etc.Vitamins. amino acids, sugars. etc.Lipids. fatty ac,dsLiposomesDNA/FNA probes

All cf the above

―― 乙亀 Z

‰j靴ゴ°百1‖][黒

AP、〈傘oい

°9

∫∫ξＣＯ「０」ＯＱｏ∽

一一０い

ごヽ■ｏ一ＣＯ夕ゝｏ

ゝ＞ｏＦ

T・ 1ご碗IヽL

ξ

∫ で

NSilllIIll[iog町)hy

Reference

Roboticprinting

Computeranalysis

rrit.ie.'fhcst al)ltiotlclle\. brrtil llou lind ill thc 1'ulttrc. tllltkc i1

sihle 1o iirllou lhc t\llrcssi()n ()f thoLlsand\ o1 genes and stuti\

blil rcgLrlatiort ()l' Itticr()biai giru th xl)d resl)onses tu ettr irtrtr

rtei cl'rang.-s.

What are biosensors and how do they detect substances?

What areas are biosensors being used in to assist in chemical

and biological m0nitoring efforts?

Describe streptavidin-biotin systems and how they work. Why

is this technique impoftant?

What is a gene array? What basic techniques are used in this

new procedure?

)pesticides

:re has been a lLlng-tenn interesi in the use olbacteria. fungi.

viruse-s as bioinsecticides and biopesticides (tatrle 42.14t

:se arc dr'firtcil as biological agerlts. such as bacteria. lungi.

t\c\. or thr-ir contponcnts. \\hich can bc used to kill a suscep

e insect. In this stction. ntajor ttses tlf bacteria. lungi. and

rses lo control populations ol.insects rvill be discussed

Reversetranscription

Bacteria

13を lcicritli tiぎ c111｀ incltlに 1ご [1ヽ :llict卜 lli β(′ (′ //``、 1ヽヽせせlC｀ ‐

1)「i1llti「 ll)

β ′/:ll′'71.ゞ

た''7ヽ お (ヽてく'/, マ/2' 1~1li、 bをlctcriulll l、 olll卜 cヽtlkl卜 lo、 ic

t()i11、 cctヽ そ1、 a、 cgctaliヽ c ccll llut cltiriilビ 、Pて

r`ultlli(l13 itiヽ 1()(luCCヽ

al〕 1lntiticclltllar i)1()tCi11モ ぃ、11l cl)、 tal thC Ptlla｀ P01‐ tll卜 :`〉 d)、 that cain

tlct a、 [il、 liCrOhitll il]、 ccticitic F()l 、PCCilic i11、 cct gr■ tip、

1:hC r(lI(1ヽlついrtiI Cr)、 itil altcl c、

1つく)｀ t'rC tO■ lkを lli:le tt)nditi():1、 in

thc llillti=tlt ilttピ l〕 lCI]1ヽ 1● rclcti、 c thcl,「 (ヽ 1い i` ll へitcr tlli、 「cを lCtヽ ヽヽit11

〔l Pl()tC tiヽ C Cnス トnlc tlnc ticti、 ct(ヽ 、ill i、 Prて (ヽlticcd(flglire 42.22, Si、

of thc ticti、 ct()ヽ in tlllit、 intcゞ I alc i1llぃ thC Pla、 11〕 ょ11lC nl卜ranc(lig―

urc 12 22わ ()(ol(■‐nl a hc、 ti=(ヽ nal―、|]をlPc(11)olc tiュ 1● tiピ h thC nlidgLlt

cel;、 a、 、11(ぃ 、、Il in llgtlrc■ 222て / Thi、 lcnd、 lo the l● 、、く,foヽ nl、 )tic‐

balallcc alld Al｀ Il(11ld inall)lo ccH l)i、 1、

Thc nlost rcccllt adヽlallccs in Our ulldcrstandillg tDfお`7(′

〃ιィ∫

′′7“ ′マ′′gノど,7,′ hゞa、 c in、 ol、icci tllc crcation of pest― rcsistant plant、

Thc llrst stcp in thisヽ olkヽvas to inscl■ thC tOヽ 11l gcnc l■ to f (こザ′

This、 ork sh()ヽ cヽd thal tllc c「卜、ttll IDl(■ cin cotlld bc c、 Prc、 、Cd in

tli30thCr()rgarli、 111[1,ld that thc to、 in、、(1、 cffcctil c Thiヽ ■■110r

、ciclltii｀ ic tidヽ tl I]CC ｀`tlヽ 1()llい ヽヽcd i11 19 「ヽ 卜) lhc p:.｀ tit:cti、 I]()f

tonlato plant、 thatご (ヽ IItiincci t11ビ tい 、11l gcllc

B r/7″ ′

'′

7ド た'′ :「ヽヽ ctin bcピ 1(,ヽ 1ヽl il〕 ic「 1lncr]tcrヽ ｀`卜ごFi tilc ccll、

1卜 、e thc sp()rcs tllld Ciゝ 、(al、 ctrc rclctl、 c(l illt()thc incdiunl Thc

Chapter 42 lndustrial Microbiology and Biotechnology 42.26

Test 師ヽ…∞メ凛tミ 遷 纂

奪 たPCB anrplificationpunJ;cation

3

亀:息 :1‐ 3?〔

:,=R=I=二 i

Tt:e Use ofOlder Tech

Bacteria, Viruses, and Fungi As Bioinsecticides: Anwith New Applications

Bactena

Viru-ses

Fungi

Bacillus thuringiensis and BaciLlus popilLiae are the two rna-jor hactena of interest. Bacrllus llitlrirlgietlsls is used

on a s,ide varietv of vegetable and field crops. fiuits. shade trees. and ornamentals. B. popilliae is used primarily against

Japanese beetle lal,ae. Both bacteria are considered harmless io hunrans. Pseudomonas.fluorescetts. u'hich conta;ns the

toxin-producing gene fiorn B. thuringiensis. is used on nlaize io suPPress black cutrvorms.

Three major Yirus lroups tliat do nol appear to replicaie in uirrin blooded aniinals are used: nucleil polyhedrosis l'irus(NPV). granulosis virr.rs (GV). and cr,toplasmic polvhedrosis virus ICPV). These occluded viruses are more protected

in the en\,ironnient.

Or er 500 iiifferent fungi are associated s,ith insecis. lnfcction an,J diselrse occur prirrarilt' through the inseci crtlicle.

Forrrnra_1or9ene., lr"lgheenused.Bar:rr,i,ria btssiattotuttllletarhi:iuntlni.top!iac areusedfbrcontrol of theColoradOpotatobeetle and rhe froghopper ir -su-sarcane piartations. respecliveiv. \;erriciiliun leL:ttnii antl Entontophrhora spp., have been

associateci rvith contrrtl ttf lphids in greenhouse and t-ield environmenls. Coelonrtr:es,:i chytrid. also rs beirrg u:ed ibr control ofniosquitoes.

Toxin binding to

phospho pids and

i∩ seniOn intO membrane

↓NH2 COOH

crystal

Plasmamembrane

Alkaline outcontents

Y

了 躊 鸞 り

/露 獅

側 腕 耐

◇了了鸞扮Protease

+68 l.Da oct're to.ir

--t- ..A.-/ sH ici

t\-t1-

SH

inside ce‖H20,catons

|

iigure 42.22 'Ihe f Iode of -.\ction of the Bat'illus thurinsietr.si.s Torin. (ar Rcleas. o1 the prot()\in fronr the parasprlral btldv and

,iell e: il lr)ss (ri ATP. resultrnc in ceil irttbalrrnce and lrsrs.949

Aggregation andpore formation

H.O cations

ノ

Inside cell

lnside ce‖

Toxin protein Efflux of ATP

ion channel

Outside cell

Microbial Groun lHaior Organisms and Applications

42.28

990 Chapter 42 1ndtlstrial'ν4icrobiO10gy and BiOtechno10gy

p°

電it胤:曜よ」:∵習滞llsPθttltlc｀ uscd Ю COmbtt tC

思模「蹴ittll薫IW[1聯lW撤猛瀑}‖

躍Ⅷittti:嘲 1∬漱1∬]i小鋤dと、

Viruses

鮒鮒III(llttlulぎ :ヽ(1撃ボ∬lCPVヽ alC tinc lCa、 t hOSt Spccillc tiltlscS,[｀

ffccting(1卜 out 200 dil｀

fcrcllt typcs()f inscctヽ ´ヽ1111■11)01ttllll C0111lnCrCiを lヽ 1ヽ1(li pCヽ tiCidC

i、 11larkctCd tllldCr thC trを ldC llalnlc Elcal fOr co11(rol ol｀t1lc cOtt011

b()11ヽVOrnl〃て'″`,r力

′、こ`て

′

(Dnc ol｀ thC ll10ヽ l Cヽ Cll■ 1=〔 1(1｀ そl:lCCヽ 11〕 、ol、 しヽ thc tlヽ C ●1 いtic―

t1lo、:iru、 cヽ thを,t htlヽ c bCCll gcllビ 〔iCtlllヽ 1110dilic(110 PI('(lt,cc ti lつ

て'lCnt

淋 ‖ ギiltttだ ]罵 illll懺il書li繊 |ギ麟 illl

thc l cCO11lbillを lnt bttctll()、 iltlヽ Prc

t()、 ill it tictヽ 1110rC rapi(11ゝ 'thtlll thC l)tl lじllt｀ lrtlヽ 、a11(1 letli tit111■ 1=Clぃ

inscctヽ 1ヽ lllarkcdly dCC,Cascd (|・‐1・ ■11、 ■iヽ ttl ■■t｀ 11 ・

‐ 1

Fungi

Funtiホ o Can bC uscd Ю COnは'in、

cCt PC｀ ｀「

ungalい 1● in、 cdl―

ci(lc、 をlS ll、 lcd ill tablC 42 11、 arC i′ ilndillg lncrCa、 111=tlヽ Cl]n tl=ll~

劇 Imで nedcヽdOpmo■ o「 boPcゞ にdC、 ム PO,C｀ い1腱Ⅲ

dゝ

Aヽ そlilalぅ lc biOill、 cctiCidCs Vヽhich ttrC tlCriヽ Cd llヽ'111 ltingl l11-

cludc ktlヽ tlganlyCillを 11ヽ (lthC l)ol、 0ヽ 11]ヽ t in titlClitioll 1ヽ)CCiを1l111lCro―

bio10=iCJ nndaboliに ヽヽuCh a、 hikkOm、cin and由 CヽⅢnぃ、卜n、 mで

actlVC agaHlst il〕 SCCt｀

The use of nlicroorgall56l-s itl industrial nlicrobiologl' anti

biotechnolo'r,. as rltscussed irl this chaptcr' does ntlt take place in

air elhical ltnd ecological r acuutlt Decisitlils to illlrke a parlicular

product. ,,,,.:1 tilso thc nlethtxjs used' ciin hale iong-teil"-',11:l-

i.,-t -un.r.p..rerj

elTects' as u'ith the apllerir-ance Llf eniiblotlc-

rcsistattl Pathogells lrountl the rr orlcl'

It{rclol.'io}ogv is a ciltical p'irr' of the area of indu:trial ecologl '

concenred *irh trackrng the tlor^'' of elL'nlt'nts atld col'tlpouncis

thoLrgh tht nirturili an't slcial r"orlds' or thc bii;sphere lirld ihe an-

t.l.rrosphcre. \licrobiologr" e-'peciallv ll\ an rtPplied discipline'

siroLrlrl bc .{)rl:irlere(l " itli'l itt stlpporting tt,g111l i"-trrld , :.-

\1iat ()r)rg"illisttls liave bce ll of itnt-ne ll-se berlel-it to llutl-tlttlttr

rl.iri,Lrgh rhc-ir rolc in riiorl prodLrction anj proccssilt 1l: :):l"'

their ir,,.iLrctt to itllpror e hLt'r'lrn lilil anirllal health' in iisrlc''rlttire'

artrl l'Lrt ril.' Iltainte nlttlct- ltrltl itltprilrelnettt tlf ellr ironntcrltal qtlal-

irr. ()tit.'r lttrarr)(rl!ii1ll\lll\' il(i\"c"L'r' ai'c ittiptltlltrlt pathilgetls llrltl

:..i.;: ,;, .1.,',i,,g.' 't'tt' rtlicr.t'r.krgisls hrr'e i.icipcct colltt'irl .r linlit

th.' .teti" liii> ol thc>c- ilartlrliLl tlltctiltlrganlstls''iht {i5i'ilra'1 r lnrl

rr.r. rrl hitri'iiitltl tlllcrilhrlii 1lr'tducl:' sltell as illllihi()iics' hlirc- cotl

trjl.irtcti t,, r, .lciribltlls ol thc hlrtllatl lil'e :lllirl in tile lasl 'clllltl'-\"

..\ ttttct,,lrtLrlrlgtrl ullt' 'iilt-ks ill lttit tlt tht:'' ltrelLs tll'brtl"t'cil-

ilrll(l!\ \iltrrll'1 'tltl:itlt'l 1ll" i'rn'lel tt-f'lt "'tltt'ttt o1'potsiilic tech-

nicll .ieir:i'.ttls"{n trccllcrlt lllil()(itl'tl(ln t() lllc lelationship

he t*ettt tc.lrn{)l{rg} arltl ptrs:ihle sociciltl trl1plicts it Sit.t:'.]t.-'

Satrtr.tcl Fltlrtlllttl t.st't'ttltitr:ntlit l'r' ()Lir-f irst challcrlge' a\ llllcl()-

[]qrl1rg111r. is to titldcr"""t1' tt-t lllLtch lts is ptrssible' the potentiltl

lnlllxtl; ,r1'tlar" lltoiiLl't '\ illl(l P1()ce\\'\ l'rll thc Ittrr'lticl 'r'tit't\ 't:

rre l1;i: \rn iitiar()bi(ril,gt' .i" e ssclllial plll-t oi Lilrs re sprrllsibiliir is

1o h.- .rble ,,' att""t'u'ii*tltte cl'fecttr clrl n ith ihe r ltriilris 'socictll

stekelrrrirrir:.. ltirrr11r the iararetriric urtl rr,rgcr'-tc* rrotc-lrtiril ittl-

f ,,.,. ,,t rrlicrtrbirrl-baseil (rillti trthet ) tcchlltllogie t'

lMPAσS OF ArliCROBIALBIOTECHNOLOGY

讚 mm“

“

ぉ bd∬ edd“ゞ

Briefly describe hOヽヘノthe θaθ 〃′」S r力υr′βθ′θ17S'S toXin klllS

are being used to attempt to conirol in

name for one of these Products?

・踪『謡紫[11=£撃[¶事部照lroadest possible impacts in your discussion of this

Define industrial ecologv'

What are the biosphere and anthrosphere? Why might you

i古 1薫 tiビ term antlЮsphere was COlned?

a particular

terms of the

problem.1

2

3

4.

insects.

What tYPes '

sects? What

Which fungi

of viruses

is a tradepresentlY

2

3

are being used as broPesticides?

42.1 Choosing Microorganisms for lndustrial

MicrobiologY and BiotechnologY

a. Industnal nlicrobioiogr tt;ts hcen LlrL'!i lril ill:ii1Ll

facture such products as anlihirrtl" irllu:1!r

acids. and organic :rcids and has had Illlrl\ ln'l

pc,nunt p,'trtite eflects cltt anin.ial allrl hurllrin

healrh. \Ii'st u'ork in this area has btt-n currietl

out using micrtrorgantrins i'olaled tlom n:llui'

or nlo,iitied hr the u:e rri e lus'il lnutation l"h

niques Brotechnologr inr'rl''e: ihe use of mo-

lecular techoiques rtl trt'ditr lnd imprtlr:

m lL I oUrg -lni \ll I \

h, Finding neu nlicrtrorsanisms in nature lbr use in

lio,".linolngl is a continuing challenge For

nost en\ ironlnalli\. onl\ a ren small part ol th

rrbser' rhle mtcrrlt'ial irrrnmunitr has teen grou'

rtables J2-1 lnd J2'lr' bur malor adranc'-.s t

trtru ing "uniuiiuretl nlitrot'e: are bting nlade

a. i"leation an,l mulatltrn crrntinue to L'e imgrnar

approaches 1-or identi f1 ing nerl m icroorgantsm

Ttrese o etl -.stahlisheil procedures are now bein

4229

conrplertlenlc.l br nloleculltr tcchniqLres. inclLrd

rng ntetabrtlic ettgincet ittg and conlbinatorirl bi

ologl. \\'ith contbinatori:il biologl ltable'12.31.

ir i-. possible 1o tr.in\iet gL'rlr's frolll r)ne t)rganislll

to rlnother oiganism. lnd to lilrnl ne\\'pr()ducts

Li. Sile-directed tlluta!enesis artd prrotein etlgineer

in! arc uscri to rttodill gctte erprr'ssiirn. Tilese

approrrthcs arc' lcadirlg lir lla\\ .tlld L;l-tcn diller-

enl plo.lLtcts riitlt tleir Propcltics l[igure {2.3t'

c. Nalural gartelic (-n-sin.!-r.lnc. is ol inctertsing in-

r.ras1. Thi! i;tr0lres erplCitilll. rlltcrobial re-

ip()n-\c\ l0 slrcrr in lldaptirc nlutitlion arld torccii

!'\olutioir. uith tltr: hope ol idtntilr ing Itticloor

t.!r j.lll \\ rllr L r ir,'t cttr, .

42.2 Microorganism Growth in ControlledE nvironments

Ir \licro,rtgalirmr elitt i.c lrirl n in eonllrl]ltd enr t

1\)nnltnl\ o1 r llitlus t,, pc: :sin-L lirlllglli.:rs lttltl

rrlhtr eLtilttrc' \)'lL'Jr\ ll L['llncr] ctrtlslilttcnts ltle

u.cd. !r()\\th pillllllL'trls alrn be rh(lscll rnd \r jc-d

II tht crrur.e o1'Ir(r\\jllr lt Illicro(llgrttisnl. Thi'

:ririrrr)irail ir Lrt,:ri l,erlrclrllLtlJ lirl thc i-tlllti.liorl rll'

trl,,. l ',:' .r':l rl I:)r-l i'

42.3 Major Products of lndustrialMicrobiologY

:r. \ r iLlt \.ttitl\ ('i..111Ptlttrlrls rle lll'lrlllc.:iJ itl rn-

tjLt.ttirLl rttierohr(rl\)g\ lllllt itllllltcl llttr Iirr'. itl

nrirlr\ \\il\ \ ltable {2'9). Thcrc irriiutlt ltltlbr

,'iL\. alllill(j ll!l!l\. olI:llli. lci(l\. hi(rlloL\rll'l\

\ri.h J\ iitc crelLrrlcrtlitt:. rllLi hio\lilli!aLlllts

rtigures "I2.8. -12.9. rrld J:.1 1) \1icr()olglllll5lll\

rlro crn bc tlscd 3\ hi(rcJlill\\ts t1l cilll\ oilt sl)c

c j1 ic chcttt tcrl 1 !'i].iilrrls r ti gure tl2. I 2 I

,q ;. "1.;;,11'.i,.' .,'1'11', 11r,1 . ,t itt'

Irlr'1ltnl jlilll,li illrlustii:tl 111j;1o[]lrltlgV atld

biotcchi〕 ol●

=ゝ

' ThCSC incltidcヽ ●idcl)' tl、 cd anti―

ttllnor agcnts(table 42.11)

42 4 Microbial Growth in Complex Natural

Environments

a 4ヽicr()orgalli、 11l gro、,tll iI]colnPIC｀ 1](ltural cn、 i

l● ni〕 ,ciltS StiCh tiヽ oヽil、 ti:l llヽ l atClヽ iヽ 11()t tlヽ Cd t()

ciCtiC n3i(｀ l()bial Pro(ltlct、 卜tli to ctlil).Out crlヽ :

「()ni]〕 cntal l)〕 arlagc]〕 ]cnl prOcc、 、cヽ llnCitltlin=

bi()rcnlcdiation Pl`int inoculatioll and()thcl lc―

1(ltcd t,cti、 itic、 11〕 thcsc ctl、 cS tilC nlicrobc、

thclnl、 clヽ.c、 :liC 11()ti'il)aI P!(,dtict、

b Bi(,dCg「 ad(ltio11:s a clitictll Part t)fl■ lttH al、 ゝs―

tcnl、 nlcdiatcd lt、「=cly t,)nlicl o●

「=arli｀

11ls Thi、

C til]inヽ 01、 c nlino「 cha11=c｀ i11を11■ )ヽICCulc rra=´

Incnttltion or nlincializatit,n trlgurc 42 13)

c Bi()dcgratlation ctin bc inl]ticllccd b).inl a n、 i tiC―

t()iヽ inClし 1(111lg o、 )=cl1 1)iCSCl]CC()l ・lい 、cincc htl―

11lic aci(1、 alld ilnc p:c、 clヽ CC 01' I ctiて 1ll) t、 tiblc

organic illattcI Rctlutti、 c dc1lalぃまcnati()ll Pl()

cccは、 hc、 t tlllticl all(tci olpi(: conditi()il、 allti thc

PrC｀ Cncc● 1● l=aniC lユ 1[ittcr Ctlil facilitatc rll()て li「 i―

ct,11()11ぃ |「CCJc tlュ ぅtC():う ,P(ヽ 1l l(1、 il¬ lt lう rぃ CC｀ ｀

ぃi cぃ 1]lcttibい iヽ l il

d Thcヽ tructurc t,1ぃ「fanitこ )`nlP()und、 inllucncc、

dc gitt(ltiti()1]11ビ Ⅲl、 titticiill コIC ilヽ 1)ccilic l● cti―

11)i〕 、()n a 111い 1ビ culc ti、 in lhc′ 11(■ 11〕 )｀ 11()Il l fl=―

1lre 42 141()i il 、alic(1 、1l titttiI[1: :ヽ 1 1llci、 al C

PiCSClう 1 こIc=I atiatiⅢ l, caiュ 1っ c t,「itご :せ て1

C L)C=]tidtiti(ヽ ぃ |]■ l n tl=C■ c ii ctil' 卜c t arlictiい tlt i11

1,lacc 入ヽl〕 cthci tl]、 卜ch=c nl:「 1:1ビ ぃ|、1ぅ

i11、

、ぃil、 o「 l1lc 、ub、u;iatc tflgure 42.17, Such

itil ic、 ctllC Cll)11ヽ 日 tヽittlヽ 1'1ヽ いh C tilcぃ 、こぃ1「うtil―

tiral l,licr(、 bial co11lnl111litic、

「 IDc,「 adatiol]can lc(ld to lll(lca、 cd t()ヽ icil) i11

1]lal]、 ctlSCS Ii l)●t nlal13=cd cを li Cl'tl‖ ) 入ヽidc―

ヽ'ICtld P。

lltlti● :i ctin いCCtll lroll c()「 I()ヽ 1()n i、 a

Questions for Thought and Review 991

particular conccrn、 vith mcthanogcns and Dど ゞ“′―

力1'jわ ″jο playing important Юlcs in this proccss

g Plants can bc uscd tO Simulatc biodcgradation

proces“ S during phメ oFCmCdiation This Can in―

volvc cxtraction. fIItcHng. stabilization. and

、lolatilization oF pollutants([gure 42.18 and

table 42.12)

h 4ヽicroorganisllns can bc atldcd to cnvironincnts

that contain complex nlicrObial communitics

with grcatcr succcss if livitt or inCn mた roh,b

itats arc uscd ThCsc Can incitldc living plant sur―

1■ccs(sCCdS_ roots_ lCa、 cs)or incll matcrials

suで h as nlicroporous gia、s_R力`_r2み

i″″is an im―

portant cxainplc of a lllicnoo瑠aniSm addcd ttl a

COInPICX Cn、 :ronn、 cnt usin=a liヽ ing microhabi―

tat(thC plant rool)

42.5 3iotechnolo9ical ApplKations

a lヽicroorganisnls arc bcing uSCd in a、 vidc ral]gc

of biotcchnological applicatioQヽ stlth as hiく )、 cn―

sOrS(flgure 42_20) 4ヽicroarra)s arc uscd t()

monitor gcnc cxprc、 sion in conlplc、 systcllls

(tigure 42_21,

b BtlctCria 、iruscs and rungi cdll bc tlscd aS bi(う in―

、ecticide、 alnd b:()Pcsticidcs(tabIC 42.141 Bて ,(〃 ‐

/″、 7カ″′ル2マ′′″、′、 iヽ .tn i!111、 lヽtani bi()Pc、 tiCidC

and thc BT=Cnc h(卜 褥 cn inc(ヽ rp)ratcd i:lto co「 n

42.6 impacts of r゛ icrObia1 8:otechnolo9y

a 11ldt:stilai nlicIく )biolo=}and biく itcchnolo=)i cを in

lla、 c loll= tcr1ll and Po、 、ibl)uncxPCCtCd P()、 1-

ti、 c and ilcgati、 c ctl_・ ct、 く)n thC cn、 iroll1llC1lt_

and o:l allill,tilヽ and intllllan、 i:l]Pactcd b)'thc、 c

tcchnologics lヽ llヽ anccsin biotcchnolog)ヽ hOtlld

bc c()Insidcrcd in a b“ )ad cc(,lo=ical and、 ocictal

cて、intc、 1 hヽiCh i、 thC lk)で、1、 of illdtl、 t●al ccolo3).

:rrlaptir e rnut:ttiit:t 9(r9

jnlil !r\phtre 99()

hinirLrlr)entelion 9X'i

lrr()ailisi\ 51 t,)i9

hioJcr:r:id:liott ()J9

bioinsecticides 988

hropeiticide 9Ex-

biopoll rncr 977

bioprospecting 96'ibiortnrediatio;t 982

biosensor 986

hiosplrcre 990

hi0lranstoilrliilioll 97te hir al 9liO

conririnrLlor ial biologt' 966

crrmetabolisrn 981

cortlinurrtts lettl 9l:.tcr nlcntrtion 9l(.)

torced er olution 969

sene rrrt\ 986

industrial ecolog-r' 990

land farming 9E0

lvophiiization 96!rrrr,/r etlect 9ii0

iltetrhLrlic c(rltttol angin!'!'rln! q6--

rlcliib()lic lllth\\ ii\ etlg jtl.':i i illi \1['E I 96r

inieroart rr lechnologr 9s6

Irrt. .,l .rl Lrr.\",ttttJrr,'t"''inirtural atlenualion 93:nrrtural genetic etrginecrinl 969

non-Nervlr-rnian [-.rotlt 97 I

path\\,avxrchitecture 967

phrtorenrediation 9tl3

prinlxrvnielaholite 97,1

proteincn!ineering 9{.6

protopia\l lusion 96-5

rer:rrlcitrancc' 979

reductiredehalogenation 919

regulat()r\ mulanl 976

scaleup 97 I

secondarl rnelabclite 97-l

\enris\,nthetic penicillin 975

site-directed mutasene\is 966

″ ´Hor,, i i it otr:sible lo creete a niche rrr rnicrohahitat

Iilr J Inirrrx)lr'iLn,rill'\\h3{ Jre mc \pc(ul F}inltof concem in tq ing to maLi .ure the microhe can

linci its best piace tc sun ir e ard function lflqrrr miehl the prr'tscnrrmic" era difler fronr

the "senontic era'- l

l. \\-hat further tcchnologicai approacbes u i11 be

requireti to culture n.:icrcrorganistlts that have

nr)t \ et been groB n I Consider the roles of nu-

trient fluxes. comntunication molecules. and

competition.

\\'het nlikc's the rrcri tr1 n:Lutltl seilL'ila !'n!1

nr'er in{ Lrniqlre ) Isn t this :illlpir ri hrl h:: ftct:

:nl t,l , It i t ,r.rlritr .. ,.1 flr. Illlr. llri.'." -: ::

sisrr: $'cre tlrst aLrle to lLrnction l\\'hat are the advanteges o1-microart:ir: for ihe

\tudv ol sene cxpresrion in compler organislli: l

|し ″‐

中議

口Ю

Ｏ

（じ

♂―″

Ｌ　貞質Ｖ

面螂　請

:ndLIStFia:

o!ogytechnology

Concepts. lu4icroorganisms are used in inCustrial micrcbiology and

biotechnologi' to create a wide variety of pr-oducts anC toassist in maintainrng and improving the environment

. Most work In industrial microbiology has been cai'ried out

using microorganisms isolatdd fi'om nature or modifiedthrough muiations. ln biotechnology, microcrganisms withspecific genetic characteristics Lan be constructed to meet

cesrred oblectives.

" i\4ost microorganisms have not been gro'"^/n or- described A

rnator chalienge in biotechnoiogy is to be abre to grow and

cl-rara.terlze -ihese observed but uncultured microorgan-isms rn ',",,hat is called "bioprospectit'r9."

. Forcecl e';oLuiion and adaptive mutatlons now are parl ofbrotechno ogy. These .:L De carried out in processes termeC" natural genai-i- engineerlng."

. The developmen' of meciia and specif ic conditions for theqro'.vth of microorganisms is a large part of industrial mi-

crobiology and biotechnology Mi-

一一■

crocrganisnrs can be grown in

controlJed environments rrith spe-

cific ltrnrtations to nraxinrize thesynihesis of desired Products.

. Microbial gro'',r'th in soils, waters,and other environments, whereconrplex nricrobial cornmunities al

ready are present, cannot be com-pletely contrclled, and it is not possible

to preosely define ltmititrg factot's orphysical condiiions.

. Microbial gr-or,'.th in ccntrolledenvironments is exPensir''e; it is

used to synthesrze high value microbial metabo-lites and other products for use in animal and

Biodegradation often can be facilitated by

changing environmental conditions.

Polychlorinated biphenyls (PCBs) are

u'iiiespread in<justrial conianlifl ants that

accunrulate in anaerobic river nluds- Although

reductir e dechlorinatirrn occurs under these

:.rnaL-r()hic cr.rndili(rns. ()\\ !€n is rcquired to

.()mpleie the dcr:radatitlrt process. In this

erperinrenl. nruds are being acrated to allou'

the final biodegradaticrn steps 1cl occur'

一ネ一

一．一、

一一一一一一ヽ〓Ｆ・〓

ギ・ [1警ξ手

||ギ 11

′「

ｉ一．・．■一

′れ

・壼．●デ鷺

）世ξｉ

ｌで■ダ

あ と 脅 〃

96,4 Chapter 42 lndustrial Microbiology and Biotechnology

human health. ln comparison, microbial growth in naturalenvironments usually does not involve the creation of spe-cific microbial products; microorganisms are used tc carryout lower-value environmental and agriculture-relatedprocesses.ln controlled growth systems, different products are synthesized during growth and after growth is completed.Most antibiotics are produced after the completion of ac

-: .' ,. ;.tive growth.Antibioiics and other micrcbral products continue toconti-ibute to anrmal and human welfare. l.lewer prod-ucis include anticancer Crugs. Comb!ratorial bioiogy is

making ii pcssible to produce hybrio antibiotics rnrithunique properties.The prociucts of industrrai microbrology rmpact ali aspectsof our Iives. These often are bulk chemicals that are used as

fooC supplements and acidifying agents. Other productsare used as biosurfactants and emulsificr-s in a r,,ide varietycf applicat;orrsDegradation is critical for understanding rnicrobial contri-butions to natural environments. The cherriicalstructure ofsubstrates and microbial conrmunity char.-.cter strcs piay rrn-portant roles in determining the fate ol chemicals. An;ero-bic degradation processes are rnrportant for the initialmocji{icatron of nrany conrpourds, especiaily thcse r.vitlchlorine and other halogenated functions Degradatroncan produce sirnpler or modified 6,,111,r6snds th;i rnay notbe less toxii than the original compoundSirseniors are unCergcing rapid de;,eloprrent, which'rlimited only by the advances that are occurring in rrolecular biology and other areas of sr ience. lt is nor,n, possible, es-pecially with streptavidin biotin-linked systerrs, to haveessentially real-tinre detectron o{ important pathogens.Gene arrays, based on recombinant Dl'JA technoiogy, allo,,vgene expJession to be rnonitored. These systenrs are berngused in the analysis of corrplex nricrobraisystems.Barteria, fungi, and viruses are increasingly employed as

bicpesticides, thus reducing dependence on chemicalpesiicides.Application of microorganisms and their technoiogy hasbotir positive and negative aspects. Possible broader impacts of applications of industrial rnicrobioloqy and brotech-nologl, should be considered in the application of thrsrapidiy expanding area.

'l'ltt nti<'robe will lnve

ndr-rstrial microbiologr and biotechrolr)cv both in-

" olt e the Llse tll- illcr,rr)r'Slni\nr\ ii) ir.hieve spccific

gotll、 、hctincr crctiting nc、、 prぃてluご 1、 、、ith lll(ヽ llclttrヽ aヽltlc or

illlprぃ 、:1lt thc cil、 irl‐ nillci1l IIntitト モ:itll l)liご lぃ 卜:ぃ 1● ■、 ti、 11 1,ti、

:itiditi● I]till卜 dc、 clて 、pcd focと ,、 ビ、、)I]I]iい dt'こ 1、 、tiChよ ヽP11ま ::ミ ltiCC t]―

[iぜ tll こnl1 11lcdical connPoし lnd、 (ar,Iibit)liご 、 11て「ヽnl ollcヽ 1l an、 一

fo「 :rlcd stcroids, solヽ iCnts organic acids.chcnlical lbcdstock、

422

anrino:rcids. anci enzrnles tilrit harc direct economic value. Thenricroorganisnrs cnrplorcd bv indLrstry hrre been isolated frorrnalLlre. end in rrurnr clr\c\. \\crc rrrodified Lrsins classic nrulation-sclcttion pr oceclurcs.

Ilioteehnolog', hu: l Iong historl. rnvr-rlving ti.ie irsc of liling or-ulrrrisnr: 1'oi' conrnrer-ciiil hcncf-i1. In thc lasl \,'\ eral decacles. the

tL'rnr hlr\ i-lacr-rnre usstlciltted u ith tire trOcliiicltiitrn ()l' ltliLror)r

sairisms thloirgir tht- usc o1'ntoleculur hiologr. including the use

irl recornirinlLnt D\,\ Iechn()l()g\ l.:ta <lrultttr l+). It is no\\'pos-:ihic to nlinipLrllrtc r.nL-tic inf'ornlrlion rrntl rlesign Ilroducts such

as pro{e ins. or Lo nrorlilr rnicr-obial gcnc e\prcs\ion. In iicltlilion.gcnciic inl'or-rnlilion e lin be ir-rrnsicIred b.'tu e eir nrerkedlr difler-e nl !r-oup\ ril'or'!lnirnrs. r,uch lrr hcluecn llictt'r-ia anil plants.

l-irc us.: ot' nricroorglinisnrs in industriai nrir:irlbii'rlir,cr, and

biotr'e hntrlogi lollrrn s li logiclil \cLiLre nca. Ii is irecessarv first torrltntrir ()r' ! r'tlrlrt lt nticroorgllti:nr thlil clilric: out lire clesir-cti

[]r'1)LL'\\ in tire rrrosl tll'ieient rlllinrcr.l-his nric|o,rrgenisi| iiren isii\cil. cither in.i coirtlollcd r-nvir{)nnlcrrt:rich lrs rr i'ernrentrrr-oritt c,rntplcx niitullll \,\ \tL'nls. sttclt lis in srtils or \\ ll[r'rs. 10 lchiu\ e

.irtiilic Srri j..

CH00SiNG A/1iCROORGANiSMSFORINDUSTR:AL MiCROBIOLOGYAND B10TECHNOLOGY

「 llc iir、 (ta、 kl()l(tn i11(l tl、 tritti l1licl obiol()ti、 1 1、 1()llin(1:1、 Lli t(lblc

11licr()o「 tani、 11l olnC that is tc1lclictlH、 ‐ 、lti卜 lc ctt、 、 to l〕 1[lilltt、 ill

(lndゞ |● ｀` tinCiヽ、cll、 tlitc(11()l c、 (l tt(■ it)nt,1 、cIっ alを 1li、 )1〕 ()十 (lc、 1lc(1

1)|()dtiCl、 ,へ 、、iclc 、r tl「ict卜 ol tll(Crl〕 ati、 c al)l)lt)を iC1lC、 (Hct,ヽ ailabic

I[11〕 ■int fr()n、 i、 ()l■ tiini n3ic,,()(01■ tlll iヽ 111、 1'1(ヽ 11l ihC C11、 irolllllCnt t(う

u、 inぎ 、●1っIni、 tictttc(1 121()lccuLtr tccil:li(luc、 t()l11()(11「). tlll c、 iヽ ling

nnlcI()()1 2tti,lヽ 1〕 ]

Finding Lttici′ ooroattiSms tn ttatttre

tinlll rclati、 cI卜 「cccnti卜 thc 11ltl(1、 ()urcc、 、)F nlicI()bittl culturcs

lol‐ t卜 c i1l i1ldtl、 trial 11licI(♪ biol()t、 、、CrC llttttlltll 11〕 11lcl iclis、 uch tt、

、oil 、ttinlPlc、 、、latcr、 そ11ld、 P(,ilcd卜「c tl(l a11(111‐ tli((｀ tllturc、 1l ol“

aH tllca、 of thc、、orld contillしlc lo bc cヽ そinlincd lo idclltilヽ 1ヽ 11[1111ヽ

、、itll clc、 il t、 1ぅ lc charactcli、 tics llntcrc、 t in huntint f()r nc、 1111-

croolga,11、 1111、 、ol biol)rospcctillg,colltilluc、 loda)

Bccau、 c olll)a lllinor Portion()f the nlicrobial spccics csti―

n]atcd to cxist il〕 11loSt CllViI()nnlcnts has bccll is()latcd or culturcd

(table 42.1)tilclc is tl continuillg cffol■ thl()ughotlt thc、vorld to

gro、、 lincsc しlnctllttlrcd・ 11l icl()o「 gtlnis11ls 1ln tCrllls of thc llll101-

c11、 ir()nnlcnt、 11l o、 i nliClobc、 that can bc obscr、 cd ha、 .cn()t bccil

culitl「 c(1(,「 icicntilヽ c(1(table 42.2, ヽヽ:ith i11ビ

l ctlヽ CCl ill(crc、 t in llll

cIぃ いitil di、 cl、 1[〉 Iュ liご「()卜 itll cc()log) an(l c、 Pccia‖ ) i11 1lnicrooI

gani、 1ぃ 、 1l olll cヽ tl C 11lc c ilヽ i10n:1lcnt、 {Tcchniques &rゝ pplications 42.1). 1llicI()biologist、 arc lcarning hO、 lo gro、ハ「

964 Chapter 42 lndustrial Microbiology and Biotechnology

human health in comparisOn′ microbial groぃノth in naturalenvironments usua‖ y does not involve the creation Of spe―

cific microbial productsi nnicroorganisms are used tO carry

out lo、ヘノer Value environmental and a9riculture― relatedprOcesses

O in contro‖ ed groν、/th systems, diferent products are syn―

thesized during gro、へ〆th and after 9ro、へ/th iS cOnnpleted

4ヽost antibiotics are produced after the completion of ac―

tive gro、へ′th i ′ 二 ′・ Antib10tics and other naicrobial produ(ts continue to

ilよ留l:∴ 11『 :infrl」∬誂∬|:鷺 rti:ふ島ニmaking it possible to produce hybrid antibiotics ν、/ithunique properties

e The products of industrial micrρ biology impact a l aspects

of ourlives These Often are bu kと hem calsthat are used asfood supplements and acidifゞ ing agents other prOductsare used as biosurfactants and elη uls fiers in aぃノide varietyof applcat ons ヽ ′

・ De9radatiOn is crit cal for understandi∩9 nni(robial cOntri―

butions tO natural env ronrnents The chelnicalstructure of

substrates and micrOblal cOnnm_ln ty cl‐ 8raCter st cs play,fn―

pOrta∩ t rolesin detertnining the fate_lfchenn cals An3ero―

bi( de9radation prOcesses are inlportarnt for the in tial

modification of nnany colη POul:dS′ especia y thOse 、4ュ thchlo「 ∩e a∩ d other halogenated ful¬ ct Orns De9radatior¬

car]produce simpler or n¬ Od fied cゥ nI「 OundS that lTay lnOtbe less t,xic thall the ori9inal COmpOund

・ BIosc n,Ors are under9oing rapid develoll1lnent′ 、Aノ hich Fli「nited only by the advances tinat are occurr r¬ g in iη Olecu―

lar biology and other areas of science ltisinOぃy possible′ es―

pe(ia‖ y ぃノith Streptavidin― biotin inked systems′ to haveessernt a‖ y rea tinne detection Ofi:mportant pathOgens

e Gene arrays′ based on recombinant DNA techno109y′ a lo、ハy

99'C eXtrF:ユ 1911 19 be mOn,tOled These systcms are be ngused in the al‐ alys s Of cOnlplex n)icrobial systenns

・ Bacteria′ fun9i′ and viruses are irncreasin91y er〕 np10yed asbiopesticides′ thus reducing depe∩ dence Oin chenn calpesticides

・ Applicat On of mlcrOOrgan sms and tino r techno10gy hasboth pOs tive and negative aspects Possible brOader im―pacts Of app ications ofindustrial:microbiolo9y and biOtech―

1¬ olo9y shOuld be cOnsidered il‐ the app ication Of thisrapidly expanding area

422

arrrino aciils. an(l enr,\ ri)es thrit hlir r- direct econitnric l.alue. Thenricroorganisnts ernplorcd bv industn hare bcen isolated frontniltr.lie. and in nlin_r cil\as. \\ crc nrociilied using classic rnutation-srlt-'et ron procecl ures.

lJiot,-chnt;lo"r hus li long histun. lnr,c.;lr ing iite irse of lilin-e or-!iittisttts foi con.rntr'rciril hcnef-i1. In the last stvcral dccacles. thete

'nr hlrs bccr-.n)e uss.ciete d u ith tire m.rirf ication of nricroor-

qanisnts thr.ouglt the rr:e r-rl'niolecular hiolrtgy. includins the useo1 r-cconibinunt D\.\ lechnolLrgl (tt,e t ltttltrrr /4). It is no\\r pos-sjblr io nlrrriprrlute g.nelic intorntrlion rinri desi!,n products sucha\ pr(itcin\. or to tnodilr ntretlbilil gcne e\pression. in aclciitiitn.gcll.lie ittliit tttrttirtp tltp bc translc;rerj br'11ren ilarkedly clilfcr-ent grouit\ o1 orLlinisnrs. ,.uch tis hetu ce n 1:lre lcril and plants.'I-lri:,sr .i,ticr..r!u'tis,ts i, i.dLrstr.iui l,icr-,bi,Jloc_r, anclitiotte hrtrrltrli lolirin s a louicitl scqLlcnci.r. lt is ne .e:srr1 Irrst iordr'ntil'r {)r.!lairlo lr nricloor!lrnis111 111,,1 cer-r-ic: out rhe desircdllr'(rLc\\ in lhc nrosl clf icie nt illllnndr. l-his nricroorg.rnisnr iiren isu\rrrl- cilhc-r in ri r'ont|0]lccl rnVrronnrent sucir as 3 ierntenlcr 0rtttirr111|li'1 niltLiJel s,\\tclns.sLrihlsinsrtil:or\\alcrs.tlrltchicr,e.1te.if ir'Sir:rI.

CH00SiNG A/11⊂R00RGANiSA/1SFORINDUSTR:AL MiCROBIOLOGYAND BIOTECHNOLOGY

‐「 |〕 c lir、 1[を 1、 k lol(ln il〕 (i tl、 tritll illicrol)101()=i、 ( 1、 lo l｀ illdをて、tlittll)lc

ill ic「 (,o「galli、 l11 01RC thtit i、 gcllctica‖ )、 ltiblC Ca、 卜 [(D l]lai11ltlin

tinci=r(,、 `al〕 d、、cH、 tlitctl l()1 じ、〔laclion()1‐ 、cPttl 11(loll ol(lc、 1lcd

l):('CI tlCl、 :ヽ 、、1こIc 、aricl) (、 1(11(Crntlll、 c tll)lD:()を lChCヽ alc a、 1liltllDIc

ra11ビ i:1= f10111 1、 01:lti l〕 ゞ1lliCI()()1‐gani、 111、 11()111 lhc cnヽ il()[lnlcnt t()

し1、 in= 、01)lliヽ tictltcd 111()lcctlitll tcchllicluc、 10 nl()cI‖ 'ヽ all Cヽ i、 ling

nllcI()。【tanlヽ 111

Finding MicrOOrsaniSItts in ttatttre

tlntil rciati、 cI)rcccntl)lhc ml(1ヽ 凛lcビ 0`1 micRbiJ culmrc、lk:)I tl、 c in in(ltl、 モlial l〕 nicr01っ i(,loご )、 で`「C Intlltll tll lllalcl ial、 stlch aヽ

、()il、とllnIDlCヽ ヽヽla(cr、 and、 Poilcd lDlCを 1(1を 11ld ll■ li((「 tllturc、 lTonl

[Jl alcti、 (「)li thc、、01ld c01nti11し ic l()hc c、 a1llillc(itO i(lclltifv、 (1を li11、

、it11(lcsiI(1卜 lc ch(1lactcli、 tic、 Intcrc、 l in litintillЧ ll,I‐ nc、、‐1ll i

croolg:lnisi11卜 、ol bioprospectillg,cOntil〕 tl c、 10(la)

Bccatl、 cO:11)a11linor portion()f the llnicrobial specics esti―

1lnatcd to cxist in lllo、 l cnvil‐ ollnlcnts has bccn isolatcd()1‐ culturcd

(table 42.1).tllcrc is a contintlillg cllbl■ throughout tlle、 ハ「orld to

gro、、thcsc u1lcul(tll cd｀ 11lic100「 ganislll、 11l tcr11ls()f thc lll[l10r

C llヽ 110nIユ lcnt、 n]()、 l nlicrObc、 that call bc(♪ bscr、 cd lla、 .c ilol bccn

culttll cti()l iticlniliヽ c(1(tal)le 42.2, ヽヽ=ith i11ご「c tlヽ Cd intcrc、 t in ilni―

cII)1う itil di、 cr、 iぃ 11liC10卜 itll cc(、 lo=) tt11(l C、 pccia‖ 卜 i11 1〕 ]icroor―

gュ 1〕 1、 in、 「1()1〕1 ビヽ tl cnlC cn、 11‐oll11lCntS (ltchniques &

へpplications 42.1, 1〕 nicit)biOIogist、 arc lcarning ho、 lo gro、 '

′ノ

／

7功ピ777'こ ′て177ピ ″J〃 /2aν

`″ι`Za,1

ndustrial nlicrobiol()g)and hi(、 tcch:1()10ピ ゝ‐ b01h in―

L/ '.'olr e the usc of rnicroorglinisnts itr lchier e .pcciiic-!oalS. it he ther= crcliiin-q na\\ itr.oalLtLt\ \,, tth nt()netai'r, r 11111.. ,11.

improrinS ihe environment. Indu:rrial nticrobiolocr. as it hasIraditir)nallv der eloped. t-ocuses on prL)ducr\ \uch as phai-ntaceu_tical and medical compounds iantibiotics. hormones. trans-formed steroids). sol\rents. organic acids. chemical feedstocks.

423 Choosing Microorgan isms for lndustria I Microbiology and Biotechnology

ilic Archaea in Biotechnol

There is -rreat inierest jn the characteristics of archaea isoiatc-tl fronrthe outllo'rv nriring regions abovc deep hvdrothermai vents ihai re

iease rirter at 250 to 3-50"C. This rs because these hartlv organisnrscan -gro\\r at ierrpeiatuie-s as high as l2l "C. The probhnts rn sr{,u ingthese microorganisnrs in a laboratory are formii]able. Fcir examnie. tirgrou,some o1'them, it is necessan,to use spcciai culturing chanrbcrs

and other specralized equipment to rneintarn ulier rn the liquiil state

at these high temperatures.

Such rnicrrrrrrganisnrs. terrned hr,pefi hernrophiles. u ith optilnurnlrou'th temperatures of 80-C or abuve (.rr:c pp. )2-i-l).1t. eonlrLrnitunique chrlierrges in nrtrient acqursition. nrctrrbolisni. nucleic acjtirepircation. and grorvth. \.1anr oi.these a;-e inaerobls that dc-pend on

cle ntental sullir tr;- 1in ic ion as oridants. Enzt'nte stabilit;,, is critical.Scinre DN.,\ llol\i.n.rases are inherentlv stable at 140'C- q,hereas

nranl' other erlz\'rllcs i.iie stabilized in r,rYo wjth unieue therrropro-ttct3nts. \4hen thcse iinzlirnes are separated fiont their proteatant,ihev krse their unique iherrrrtrstebilitl,.

Thesr cnzrnrrs rrra., have intportani applications in mernlne prl-cluctjon. nreiai leaching and rel-over,,. and in irnrnobilizeC enzyme sys-

lrnrs. in atidrlroir. the prissibi litr til'selectivc stereochentical mtxlifi cationrtl conr-Lrltntls noiliallr not in solutiort al Ltrt'cr tempcratures inay prr)

\ i(lL- n!'\\ r(ruic: firr directed chernical sYnthcses. This is lut excitrng alde rirrLnrlinl arell !11 th. nrrxlcnt biololicai science: to nhich environnttit-tel nricrobrol0gjsl: cul.r riluke .ignitlcaltt contnLrutions.

Estimated Total and Known Speciesfrom Different Microbial Grou

E sti m atesin V;:ious

of the Percent "Cultured" MicroorganismE nviron ments

EstirrratedI'otal Specirs

Knou n

S pecies''I)erce n !

Knorr n

[-nr ironnte^1 f-sl irnatcd I)crcent Cuitured(iroup

Viru ses

.Archaea

B rLc teri:r

Fun-ei

130000b

40 01Xlト

150003060000

5000<500480069000400()0

14]ビ

1121

う

67

Seawaier

Fresh q'ater

Mesotrophic lake

Llnpolluted esiuarine s,arers

Actir,ated sludgeSediments

Soil

()001-0100

02501-1001-30

_11_15 _′

02503‐

'Mid-1990 values and sh()uld be rncrcf,s€d li) 50!7.hThese values re sub\untiail\ uildereslimnred. p€rhaps b) l-l ordr.\ oi rnaqniludc

'l I indicales fiat these yalues ee probahl\'!!ro\s overeslimatei.dSmall subunl (SSU) rI{NA dak indicxre rnu.h hrsher in \nu di!enrr} dd Si.cn h\ cukure h:rrd sludi.\

Adapledfrom:D.A Cos3n 1000 \'ri.robi!l genonr\-theuntafrprdresourr fiSrr,cli Il:lJ-l6Table l, p. 15.

illcsc Prc、 1()tl、 l卜 ttllctJitllcti 11licIぃ bc、 11]、 ,iticiil〕 ゞthc Llヽ Ct)1)ヽ i11-

glc― ccH =cl 11liCrぃ c11ご 〔lI)｀ tll(ltiぃ Il を11)IP「 いtiChC｀ lhtil llllぃ 、、 :ltill iて ,111

(11■'tlsi()1l alltl nRicrt,bitll さぃ:]lll]t‖ liCatiぃ n 、、1lilc :1〕 illinli/iln g 、)、 'cl―

gro、、ith b) 11〕 o「C rtllDill―

=「

o、、ing c、 )11]I)Ctitい I、 1 ‐ ‐「 1 11‐

Genetic Manipulation of rh/1icrOOrganisms

Gcnctic lllaniplll(lti(Dn、 arc u、cd lo producc i〕 ]icF()Orgallis11ls、 1ヽ111

nc、v and dcsirablc char[lctcristics Thc clas、 ical nlcthods of llll―

cI()bial gcnctic、 (、 どで'(/?て り,′て,′ ノマI Pl(1)tl、 ittil rt)ic in tI〕 c dc、 ビIぃ P―

11lCl11 01'ctlittlrc、 lbr irldヒ lstritiI 11〕 icrobi、 ):ぃ=、

MutationO1lcc a prollli、 illビ ごultulc i、「 しlnd [1、 Liriピ l卜 ぃi tcCi〕 n itltiき ごヽti:う 卜C

uscd for culturc illlpro、 cnncnt illciLItiillご ご1lCrllictil l■ utagcn、 [lnd

Source: D A Coq in l{I\). \4 i.r ot,ial _cenonres-the ur lapped resource. Ibrec, I 8: I 4- I 6. Tsble 2,p. l5

tlltiti、 1()lci lighiぃてて■ノた1/Pr`′ ′ ノノI A、 a llじ 、arllplc.(1lc ril、 t culttllて

ぃ1 /'で ,″ ′(′ ///″ ″77■ ,「 tl子 |″ ,1 ヽヽh i t111(■ tll(l bc tlo、、11 01Rl、 tllldCrヽ tati

C01ltliti(、 11、 )iCl(lcci l●、、 tt,I]CC1ltl tlti()11、 ol PCnicillil]In 1913

、lrain o「 /t,′ ,ltア /ル ′`′

″ (カ ハ、て,マて'′ 7'′′7'ヽ aヽS iSOlatcd― straill NRRI

1951 -― 、、1lich 、、tl、 lullhcl in]PI o、 cd thlough lnutation(「Igur

12.1, ‐「 oda)ilno、 t pcllici1lill l、 lD1 0(luccd、 、.ith et′ ガ(1′ ′′′

“′7,(力 ′]ヽι

マて'′ i!`′ ′? gi■ )ヽ、n il](lcIぃ tDiCヽ tilrcd iモ 111lcntcr、 、、hich gi、 ics 55-1'ol

highcI PclliCillil〕 卜icld、 thtln thc origillal static culturcs

Protoplast Fusion

I)rotoplast ftlsioll i、 Il()ヽ ヽ ヽヽidcl〉 t.、 cd、 ith)casts and lnold`

1ヽ● 、1 01 lhc、 cl]liご iい い1=tl I]i｀ 111、 tll C.lヽ Cヽ tltll()r()f a sirlglc lllatinl

モ)1・せ ｀`hiCI(1ご こ:き ょ、きヽ 1:]ビ ごl‐ttilぜ こいi l ttinti()111 11ltlttltioll、 lh〔it coul(

lct`」 1(1、 ti t:ill tit=さ ilcl ti:い い Iて こヽa rl、 く,tit tcnctic、 tudic、 ヽヽith thcsl

1111ご [、 1● Iピ ti「 i、 111、 1■

1 1ヽ1 li〕 i tt、 1、 tii C PrCPtirCd b)i gro、 ving tllc ccll、 il

an ist)lonic、 oltlllon、 hilc trcatinゞ thcl11、 ‐ヽith cnzylllcs includinl

16 Chapter 42 lndustriai lr4icrobiology and Biotechnology

NRRL 1951[1201

1

NRRL1951 B 25[250]

|ド

X-1612[500]

|と・wis Q176〔 900]

|:

Bし3-D10

Pt:nit'iliiunt rrtqtte.fttrrii have been l'used r"'ith those of P' cltrt'so-

qettiun. Even veast proitlplast-r and ervthroc\'{es can Lre fused'

Insertion of Short DNA Sequences

Sho|t itngths ol chcnricallY si'tlihe siz':tl I)\'\ seqLrcnces catl be

inse rlcd intii re cipicnt qlicro(lrgenl.5111s hr tlle lltirce ss o1' stte-

directecl nrutagenesis. l'his can crciitc \lllllll gcilcllc ltltel lltign.''

lcaciinglitachiurgr\)1'Olle()iscrellt] atllitloltcirlsil)lhcIal('L'tplo

te in. Sirch ntin0r antilto liciC chrngcs itare br'rll lilLrtl'-1 itl lclld- in

lnan\ cuscs. to uile \Pcctcii ciletlgcs iil prolt'l;l chlrr:ritcristics' antj

itar.e t.esulietl in nc$ prodLlcts -sucit :i:; Irlilie cilr it'trnttlctllltllr re

sisl:iltt erlz) tllels atlti e llz\ lllr-\ thlit clin clitltir zc tle srred reactiot]s'-l-he'c apprortches ltrc ilal.t o1 tilt f ie 1tl o1' protein engineering'

49133P,23q

48-1655

49-482

49-2695

60-529

50-1247[1.506〕

48-786

|

i tl

48-1372[1,3431

1 ‐i‐

49-901

49-24291 Fi

50-724

50-1583

1 :'1,

51-825

1‐52-85

11■ |

52-817

11■53-174

:|‐ ||

53-844〔1,846]

llc,r.,are industrial microbiology and biotechnology sinrilar and

different?What approach is being used to increase the recovery of pre-

viously "uncultured" micrcorganisms from the environ'

i'n e nt7

What rs protopiast fusion and what iypes cf microorganisms

orr UsPi ir Inis orcr.::lDesci'ibe site-directeci mutagenesls and how i1 is us:d in

biotechnoiogy.

!!lat is protein engineerino?

3

)-2166

50-25

0-935

茸＼、

53-399 53-414

12,658] [2.580〕

馴fm ttf∞

馴翼

F

51-20A2 51-2033 F3121140]

F3‐ 64

12.4931

'igure 42.1 \Iutation \I:rkes It Possible to Increase:'ernrentali<)IrYieltls. A"gcncalogr'oi'thcttttltlLtirrlll]1 |LL'\\L'\

rsetl i0 inurcasc pc-nicilltn Iieltls $ith PdtitilliLtttl (l1t\'\()!(tltttlt

tsi.g X-t:uy,treilt.te.t (X). U\r trcat,iL-nt (Llyt. allcl llltlstard Sll\

N t. BY use 01,thesc lllulatioltal processes. the ficlci lr as itlcreas.'.1

l.ont 120 lnternarional L.lnits (lUtto 2.-580 iLl. a l0-1'old itlcrease .

-.innrarketl transfers u'ere usecl for nlutallt cro\\'th alld isolation.

fields in international units/mi in brackets'

_.ellullsc and heta galiictr,rrr,nidase. The p!otoplasts are therl rt'-

rtncr-atetl using osrnotic stabilizers sttch as sLlcrose Alter rcge n-

.:r-ririrrtt rtf the ccil ri all. rhe neu Prr:ttlplaslll 1'ttsitln prodttcl clltl tte

ir'eJ in furthcr studies.

A rIa.lrtr adr lntage ()1' the protoplast f'ustorl tecltlltclLrc i. thlt

protoplasts ot'dillcrent rlricrobial species cu be fused' er en il thcr

,ra nu, closelr lirlked tarontlllrcall.v-. For cxatrlple' protr-lpla:ts oi'

Transfer of Genetic lnformationbetween Different OrganismsNcr,, rrltcilltitrr, i.iare lrir,:n throLrgll tilc liilllslL'l ,rl nLlcle ii.lci.ls

l-..'1..'.rrtrtii1l'ercrtt (it !l1llt\ll1\. uliitll ir Irill'[ ()i tirt'rlipirllr 11t'r't'lrll'l

ing l ie1t1 Lrl ettntbinttorial hioloul t tablt -ll'-1 r l his inr trlr et tire

,r,r,.,.1a, r)t' !f It!-\ fl'ttttl tlll.' ot-latlisttt it-tto rllolhcl" gir rllg tile lt

i:iprr':tt rltitctl iellltitilitl.': .tr.li lt' ltll itltlce'cti ' l1t'ltitr t" 'lllrr,riti lrt.ltri.ltlilr,il de!tlt,l.ttl\rjl .\l llll|r)11"1 l '''ll\ c\iilllplc (11'tlli\

ap].,',rach \ a: thc rl ailil{llr (r1' th- 'stipcrbu!" paicnte tl b\

{ \1. f llikrrlribat-tr rtl l(lli. ri irich hrid itrl lllri'll\c(l caputrilih o1'

1,r,11,1.111irct1 .it!tltti;-itirltl. Th.'-lctle: lirt liiltihlotic plrltlLtcltoll call

lre lLl:l:1'L:l'letl irr:l itlliirrt'l-!lLilitllt l]llti lllLrtiriie::tilrrlllcrllliihi()ti!''r)1 C\tn ti) ii Iloil;11111f!ir111'-lllrl,lrieitl-r illiclt'L't-!'tltt'trl othct er

Itttlllie s ilrc tllc e\fi'\\i()n' tl !- ' "lt ()l ihL'

'll1) lllr !lt'11illrill't\'

tl-rrnr l)ir'rrrlr,rttt)tt(t.\ Pttiitltt lrird tltc ptiltlLtetirrtt o1 Pccliocill' a blc

tc'r-i0e tlt. ill a r cast ust'tl itl lr tl'tt' l'ertltelltltti0tl t" 't ihe Prtt lilre It

aor'tlr()il i n !' i.ltcteriel aolrl,tl I I I Illlllt \'D\.\ erpr.es:ioll tn tlitlcte-nI I rl--]llrll\lll\ Lilll llllPl()\t lltrlduelitltl

elllcicncr and ntinintize thc Pr-rnllu.rtir rtl rte P\ rcquiretl bctorc' the

prtltiuct is reat1l lbr r-rse , For cralllple. rectlttthtnant hltcultlr it.uses'(.\c(

lp. 10'1 5) can be repliclited in insect ia:-rae to achiere rapid

l:u s.' sctlc prrtiuclion of .L de :irctl r irui rrr pl ote i ;t. Trltnsuenic plants

r.1i,.71.r, i.,.,';'..'l', -i-':il1.r\hr'llt("11r)ll'.ilil'lr!liriilrl:j(qu'rtrl;-i 1.. ,i : ..:i .i\ ,.1 i;..:.:r..:. :'t ,.ii;.i. \lli"'' ll:l.L:;1.ll r' ''r :r' I !rl-

Llrll!ll irt '-. ';5i. f)\.\ :,li,.r l: Iri-lili i' irr 'lillllr 'll'rrrt 1l lil r'i\lil-l

D\ \ err:rtL'ri illiirt'ilt,'1:.l,la' ;iil!i il :lini'-!r'lil r\('i 'i ' 11'1i I t tl:'

.\,,rrtlr.;:tn!3 i)I ga!lal1! iill'trl'illelt,ln il15r) ai1ll ha ill5'l-i'Ll lntO

nlie fr)organisttls li:ill! \ iat(i1'\ lnd re conlbitlalll DN.{ tecllniqtir's.

47-638[9801 47-762

|

47-1564[1,357]

5120P,521]

|ド>＼、、、

52-318

' 嗜可~~¬

52-1087 1

425 Choosing MicroOrganisms fOr lndllstrialヽ 4icrobiolo9y and Bictechnolo9y

Property or Prodtict′I■ansferred

～licroorganism tised cOmbinatoriaI Pr∝ ess

Ethanol production Esc!rc rit ltia coli

Combinatorial Biologyin Other Organisrns to

1.3 Prooanedioi prociucrion L.. coti

Cephaltrsponn precursor Penicilliun tltnsoqenutnsv nthesi s

Lactic acid prrrCuitionXf iitoJ production

flrcatini n aseb

Perii oci n'

.{celone and butanolproti uct iitn

in Biotechnology: The Expression of Geneslmprove Processes and products

'\tttchurottt.tce.s t ereti-slae A muscle bovile lactate dehydrogenase gene (l-DH A).expressetJ in S. ceret,isiae..\. ceret'i:iae 957r xvlitol ccnversion from xylose u,iis obiained bt. irarsfr,rntins th; XyLI cene of plclla

rlrpilr enr'odin: a \r lose ,.J,u,,,." i",, a. , , "r,,;,,, "

,,,f ,,,, iUi. ,, .rrir,,, ,rl,],.i.,,,orgattistlt lbr the protiuction o1'rylito1. u,hich servcs as a sweetener in the tbod in6ustrr,.

E' crtli Expression of the creatininase gene fionr P.seutLtmotut.,tT;aiilri R:65. Cese inserted *,rth apt-rCl8 vechr. .,'

lntegratron tlf pyruvatc decarboxi'lase iind alcohol dchvtirosenl:c ll from Zyntomonus ttutbi! i.s.Introducrion of g:nes rrom the Klehsitllu pncutttortile ait re-gion into E. r.ctli rnltde pcssible

:rnaerobic 1.3-propanediol productiolt_Production 7-ADC rld 7-ADCA' precursors b1 incorp,ration rrf the expandase gene of

Ce phalosoyxt rin ac rentatiutrt itto penir i I I iutn by transfcx-ntarrun.

lirpression of bzrcteriocin t-ront PeLliotot.r;u.v ut itjilutriii in.!. rez,r,i.iiae to j;rhibit u,ineconlantinan ts.

Intrutjuctir'rt rrl; .hrrrrle \c\'r,,r r''1,, L.,tL.t I,t,tt',rr, /,,,. r, r,r rJrfr,,\(J (lrrrrl,lrilrrslufnr.,ironprotocoi. rvhrch resultecl i n ;crr,,nr- rnij bui.-r nLrl lL,r r l.ju!,n.

S. i ereyi.siue

C Iost ridium d( (tobutyI i( utn

一

，

守

「

ACA=?aminmephrlosfomnicacid:7,ADCA=Taminodecacetoxyccphalosporonicacid

t.t,.iue. lu:t ljio-l7 {5h \r1v'f i,JUr

Adaplcd from S ostcrsaa劇 :L01s、 Onland J Nieison 2(〕〕,MctabO:ic cnginccin8 of Sα εε′r7r77・ ●`′

∫ε“でドなた′ピ MicrObiol MOI Bi01 Rev 64(1た 34-50

\ e ctoi's l.ttt, scr'tirttr i J.-it jnclrrilr artif icial chr()ll()-\()ntL.s \ri.h asilrosc 1'or' \eltst\ (\,rCs). [rac.icr.ilL (BACsl. I)l bacleriophage,tit'i r c'd t lli'r)al.soire s (l),\cs ). a'd ,*1rlr,ilia, a.lif icial e hr,,nr()\( inlr\ (I,1.A(-r ). )'AC's ilre csl)ecilill\' \ arlLrahle because liir!cD\-.\ scqLren.c:; 1()\er l0(l ktr) (.a, Lre ,laint.inr.rl in the \.\f.ris I\eillltsle ait11rlnor0ntr tjt \ L,tiSt Ce ils. ,{ g0ttd erlr1lnle 0l 1gg1111.

Lr\. l\ Jlro\ idctl hr tire r lrus that causcs 1'oor,anr1 rlloulir r-liscascof iatilc anrl other liiestock Ge nctic jltlorrrlrticln tbr u lirot-and1l()Lrlli \1j\a t\c r irLrs untisCn calt hc incol.ir()trtled inlo l.,. r oli. l'ol-liirr ed t.r lirc r'rPreriron .i'this gcnctic infirrnmtic;n and sr.nthrsiso1'tlte .gent, Itr()(lLltl t iot i-rsc in vaccine Ilr()duction (figure .12.2 t.

(icnt-tie iitt.ritr;rlirrit tra,s1'cr all.*s thc. ltr.ducli,lt rr1. spe.i1r. llrotdin\ rinrl pr'ptide -s u ith.ut contJnrinarion bt siinilarirr.tlucr: thut nrrghr hc sr nrhesizttl in thc original or-sirnisrl). l-hisliPp,.rrrr1f1 aiilt dL.arr,.rsc lhc lintc lutd c(ist 01'rCc<trerinl and puriir rrg u Jrotlrrcl ..\nrtthcr ilta j()r t(l\ ailtitge ol'peJrlrde prodLlclionu rtir i-riotechnologr is tirrt oirl_i Lrioiogicallv acti\ c stcrc()is()rllcl-sr1r'c llr()duc.d. t his :Pcciticitv is rLrr-luired to avoid the possibleharrrlirl sirle eflecrs ()i'ir.cti\,e stcreoisonrers. as ()ccLlrrecl in thethal idontide clisasler.

What is combinatorial biology and what is the basic approachus€d in this technique? What types of major products havebeen created using combinatorial biology?Wh,v mioht one !.!/ant to insert a gene in a foreign cell and howis this done?\l/hy is tt i,nporiant to produce specific isomers of prcducts foruse in animal and human health?

Modification of Gene ExpressionIn ad,-litirirt trt inse rtrn! lc\\ ,jL-nc\ rn .rj:u1t.nts. it ltlso is ltLr:siblt to ntodilr gL,ltc iaqilllt jon br chalginr, ,,cne tr.in.ijl ll)tiirn. lu:ing llr()teins. r|r:litin!, hr br itl llr()nr()iers. lrnd re rnor in!1'ccrlbirck regullition r.()ntt ols. I hc\c liIlll()itr.llc\ ntekc it lto:.:ihic ltt or e rprodLtcc u rr ir.it r lir.iclr o1' 1trotlircr:. lis sho.,r n intable 42.-1.

l'he ,taclil'icatiiilt.1 gc,c r\Jlr-c\\i.n .rls. i;i. itc usetl [. i.icniiona]lralternrellltrolicpiith\\irrsb-t in.reirrationorder-eLulllronof'.[.1-!iljc !]unr-\. ',', hrulr is tlte f'ie ld o1' pathtar architecture. I-,n-clerstanding pathu'a! llrchitcLrlLrre rriuk:s il J)ossihlc 1O rlrsiln apalh\\'av that u'ill be ntitst ell'icic'nt hr rrrotciin_q sloriet-()r en.rg.ticallr niore costl\ routrs. -l'his arrpr.riiir hri: hecn urerl 1o ir.rpr.rcpenici J lin prorluction bv rretabolic pathu a-r €ngineering (\l pl._ r.

An illteresting cler elopntent in ntodil'r irtr Se rrr c\l)l-r\\lur.u,hich illustmtcs metabolic control rngincering. i, th;rt ot tlrcrittg c()ntrols lOt the svnthcsi\ ()1' l) copene . an jJIpr)riJitt iiltttr.,\ldant norrnnll)' present at hich ier cls in Iorratoe s anci tonrrrttrproducts. In this case. iin enqineerctl rr.gulalon circuit u a, clc

signed tit control lvcopene s\,nthe-\is in response to the inte rn.ilmetabolic state of E. coli. Another reccnt derclopntent is the u-se

ol modifled sene expression to produce varianrs oi rhe antibioricerythronrycin. Blocking specific biocherrical steps (figure 42.3)in pathu'avs for the sr nthesis ol- an :tntihiolic precursor reqr-rltedin rnoilil-ied final prrtrlucts. l-hcse rrite ; eti pr.rJLrct:. ri hi,:h hiir.-slichtl-r'ditlercnt strriclrlre\. ealt be ir,irJ irrr lheir p,r::ihlc :rt:-tinticrobial cftects. In ailditiirr. ir,i ihe Lr.c .i rhis appririi.h. ir i:p()ssible to better establish thc \rnrcture lltrctii.rn rrlatir-rnships ot'antibiotics.

VP#1protein

Foot-and-mouthdisease virus

Viral RNA

4

Viralproteins

Beversetranscription

Clone oi

ごVira DNA wlh

VP,1 9ene

謗¨I RettrctOn

l enZyme

r繹『鰤中

1

|

▼

不 :謂献

ll習∝mttOn d

Ａ

ａ‐ＲＮ円（ノ／ｔ、．ノ／

Ｖｉ

ｆ。

Foreign gene

Bacterialchror]]osorrte

ヽヽ＼＼ヽ、

VP11

protein

recomb nanttl acte ri a

Figure 42.2 Reeornllinant \accine Protit-tcti<ttl'

( r1' rer()tithi Itlltll D\.'\ 1c'cllrl tqLte:. I firrrl litltl t tlr itlih

aoaGa

30eVP#1 protein from recombinant bacteria{or use in vaccine Production

( itni. cotlinl tor ile:ircrl Jtroditcts can be e \Jrresserl in dilJ'ere ilt ()rsallisrlls. Iil' the use

.1i:,:rr.. \ 3.c jn. is prorluccd titroLrgit ciolting the vaccine gcne\ int() l:.tclterit-hia coli.

09鼈ラ

E^*rpl"r "f R".ombinant DNA systems Used to Modify Gene Expression

AcrinorhodinCellulaseRecombinant protein albumin

Heierologous protein

Enhanced gro\.. th i:il.".-\irlro aciri.:

St repirnnt t t s coeI itoIorClostridium genes in Bacrllus

S a c c h a ro t t t t c e s t- e re v i s i a e

Scccft arornr ce.i t' e ret i s i u e

,4.tpe rqi I ! u: nttlu litns(.',,t^,tt lu; t, t iiitti

Modification of gene lranscription

Arnplification of secretion through chromosomal DNA amplification

Fusiun tu a high-pruduction protein

Use of the inducible strong hybrid promoter UAS."1/CYCI

f)\'erproduction of glyceraideh-'-de-3-phosphate dehydrogenase

l:L)lxijL)r rri bio:r'nthetic Sene-\ rhat lead to enhanced enzlme :ctir ities or

r( , .Jr,'1 t:rJbr.k r.lLlJIl\\n

こゝ ostcttadrd:LO`sni mdJヽ た1%n2α"Mctab01で

cnsin∝ ing Or,ε「′口

"″)″∫εご″■5滋ξν:ε ,々夕ο′″ο′θiοι Ra■ 6“

'I)■50 Tablcl p 35

968

427 Choosing Microorganisms for lndustrial Microbioloqy and Biotechnology 969

ム

4ヽodule 2 Fvlodule 4

lヽodule l Modu e 3 lv4odule 5

Modified Structures

卜Иodule 2 4ヽodule 4 卜Иodule 6

Modu el 卜Иodule 3 4ヽodule 5X

3 ocked

einzy『 ln e

rv4odule 2 圧4odtile 4 卜4odule 6

Иヽodule l lt4odule 3 Avlodule 5

XBIockedenzyrne

，Ｓコ一】ｔ

Ｏ　ＯＳ聟ヽ「工一

Ｏ

C)thcl c、 tll1lPIC｀ illcltl(lc ti]c il]ビ rcti、 c(1、 ):]111c、 i卜 (,i tt1ltibiぃ tic、

al〕 d ccllulasc、 11,()(11lCtitiぃ 〕()1 ビc11ご じ` |)rCヽ i`t'I]l))ヽ へ al111っ lil'ictl―

11()13 grctltcr I)1()(cill )ヽ11[llC｀ 1｀ _ tin(1 11]1ご l tictiヽ C C11/)1llC ()ヽ Cllつ l｀ )(ltic

tioll o「 l c11〕 ●ヽ 111い 1 ltctibtit k i1lhilDit,ぃ :l Rcc● 1111｀ :Illlnl l)lょ 、1lli11ぃ ピc11

11)rc、 こ11111)lC 11｀ tl)(()111lD「 i｀ C ]()(o■ 1)`, ぃl thc 、、)l tl卜 Ic l)「 ぃicin ili ti

nlo(111lcd、 llaill a tc1litヽ 1(l iI]cl c(1ヽ C i11ビ ぃ1lCC1111を 11101〕 ()、 cl that ill lhc

oligint、 〕、tl‐ aill

Natural Genetic Engineering

Thc nc、vcst approtlch ilol crctlti1lg nc、 、 :1lCttib● lic capabilitics i11

a gi、′en ■licroorganisnl i、 tllc arc[l ol natural genetic engl‐

neering,、 vhich clnplo).、 frlrced e、 olution and adapti、 ie nlu‐

tations r、 どど/,″ 2∫ 9-■`', 1‐

his i、 lhc P!ぃ cc、、ol uヽ ing、 lDcc illic

cn、‐iI()innlcntal strc、 cヽヽ tい ｀「(ヽ rcc 1llicrぃ ぃ1■ tl i'iヽ nl、 1(, 111ヒ ItatC

tind をldtlPt thヒ 1、 cFCtitinピ 1111ご iて (`|「

=till i｀

子]]、 、ヽi〔 1l 1lc、、 hiい |ヽ iゞctli

CaPCthilitic、 1｀ hc 11lcピ Il till i、 In、 ●i tllビ ごヽ tidtiI)li｀ C nl tl〔 まiい ntll

P10CC｀ C`｀ incitldc Dヽ A「ぜtl lユ l11ご t i11● 1l t、 in、、hiピ ll irtin、1■

)｀ tilコ lC

clclllcnt、 and、 arious t卜 pc、 )`f rcct)1lRhirltllion Iつ i ll)critiCtil r()lc、

as、 ho、 n in tablc 42.5.

HO%′

一

.へ lthough thclc i、 llltlビ 11ご ()11[1て ヽヽCrヽ )CO1lCClllilng this arca,thc

lC｀ lD011｀ Cヽ いI 11]icl o()igtilli、 111、 1い 1ヽ1せ ヽヽ lD「

い`‐1(lc thc po(cntial()J

=CnCrtll l〕

g nlicrool=alli｀ 1ュ 〕、 、ヽith ncヽ 、 nlicr()bitil c1lpabilitics lk)「

tl、 c ill i1ltit卜 tl itl1 1]licI()IP i()1ぃ ぎ)tinti t)i()lcchl101()g).

Preservation of Microorganisms

(:)ncc ll nlicroo「 q(11l i、 11l ol 、l rtl、 1lt卜 bccn 、clcctcd or crc〔 itcd to

scr、 ca、 PcciliC purP()｀ C il intl、 t bc Prc、 cr、 cd in its original fornl

「or itll■ hcr usc and sttld、 Pcliodic tra11、 lcrs of cultllres havc bccn

t、 cd in thc ptl、 t、 altilougll thi、 can lcad(o lllutations and Phcno―

t),Pic changcs in lnlicroo「 gani、 1ln、 Tl)a、 oid thcsc Problcllls、 a va―

1lcI卜 ol culttlrc prc、 crヽ ation tcchnicltics lna)bC tiscd to lllaintain

tic、 ircdご じlttiTc chtirtictcli、 tic、 ltal〕 le 42.61 1´ }ophilization,or

r「cc∠ご_til卜 i11= tilkiヽ 1■ I tlビ ごiI' 11(ltliti llilliヽ

=cil ttiC trCqucntl).clll―

「lて いきti｀ i`tll 1llic:ぃ ●ritilliヽ i■ 、 へltht)ti=1l lぃ I`Dhiliztitiollを ln(l li(luid

ilitrて 、ごcl]｀ 1('I tlゞ C CtiC C● :い lDliCtit(:d tllnd rc_ltiil‐ CC、 Pc ll｀ 1｀ C Cquip―

nlcnt thc、 do tlHo、 、 11liC1 0bial ctilturc、 lo be storcd ft)r ycars

、ヽ1lh(ヽ Lll ll｀ ヽヽ ぃま｀、itl卜 ilit、 ()r til][lCCunlultitinil of nltltations

OO

O

lv・ odule 6

SS^- n:::--<".<O:.=

).,'rr HHo,.,. r> o, ---<HO> HO', /

', .<

" HO>i)

HO"

O

HO“

HOr,,

O

HO‖ "

HO HOJⅢ ‐

HO

一

DEB

970 Chapter 42 lndustrial Microbiology and Biotechnology 428

Natural Genetic Engineering Systems Bacteria

Genetic Engineering \{echanisms DNA Changes Mediated

Localized SOS mutagenesis

Adapted 1ia..nes hi iii n g

l-n-i.'1-n9. Tn.i 0 precise excistott

In vivo delction. inr ersion. lusion. anC duplicationftrrnration

Tll" II topoi.rn]cra5e l(cunihincll,,nSitc-specrl rc iecombinatittn (tvpc i trpoisomerases)

Trlrnrpo'ahle ricrtrents r1116r rpecir:r

Dl'NA uptake (transforrnation competence)

Base -substitutioirs. iranreshitis

i frameshil'tingRecipriral recon.ibinaiilr, oi'1lanking 8/9 bp rcpeais: ,e\iores orlgin,ll sequettc.

Cenera111, recip:ocll recombinlrlion of shorl sequei-.ce reireal\: occasionalll nonholttolrgou:,

Deletions anC fusions Lrv nonhomolotc)us iec()rllbinatjon. sometimes at shorl lepeu'rs

Inse(ions- ercisions/deletiitns. irr ersicrns bY concerterj or succcssir e cieaYage ligati(ln ieaciir)It\ Jt

\horl \(qllella. Ien. JI.: i,,icrll(. ;itt.;t.:tlchc.Insertigns, 11ansnositions. rcplicon 1lsi1ns. arijacert deietions/excisions. adiacent inr cisions br 1ig:iticin

of 3'Cll transp.r ton ends of 5' PO. grouirs llom silggered cuts at nonhonrulosou. tarscl .iLc.

Uptake o1'single stranrl indepcndrni ol se(lllcn.c. or of ri<;uble-slranded DNA carrvin.t spc, ie.identillei'sequence

Adap{ed from L A. Shapiro- 1 999. Naiu.al generic engin€ring. adaplive mutalion. and baclerial .volution. In MiL robial EL tios\ oI lnfedit)us D isedt?

for Microbjology. Derived from Table 2, pp.263-64.

E Rosenb,:rg. editor.159-75 washinglofl- D C: Americrn Sditlr

Methods Useci to Preserve Cultures of lnterest for lndustrial Microbiology and Biotechnology

■lletilod L'0mmerlts

Periodic transfer

N{ineral oiL slant

N,linimai metlirrm. clisiiIlerl qater. or'.\nter agar

Freezing in gros'th nredia

Dry uig

Frrrz--Jrr rrrl,.ropl:rirz;rl.,r, t

Ultrafieezing

Variables clperiodic transfer to new media include transfer liequency. medium used. and hoidingtemperature; this can lead to i ncreased mutation rales and production of i'ariants

A stock culture is grown on a slant and covered rvith ,rierilized mineral oil. the slant can be stored at

refri.gerator temperature

Washed cultures are stored under refrigeration: these cultures can be viable fbr 3 to 5 months or longer

Not reliable: can result in damage to microbial structures: *,ith some microorganisms. however. this can

be a ureful meanr ol cullure lnrinlenance

Cultures are dried on sterile soil (soil stocks). on sterjie filter paper disks. or in gelatin drops: these can be

stored in a desiccator a! retiigeration telrperature. or frozen to improve viability

Water is removed by sublimation. in the presence of a cryoprotectiVe agent: sealing in an arrpi:le can lead

to long-term vrability. u'ith 30 vears having been reported

Liquid nitrogen at I 96'C is used. and cultures of fastidious mrcroorganisms have been preserved ibrmore than 15 years

4.

1. What types of recombinant DNA technrques are being used to

modify gene expression in microorganisms?

Define metabolic control engineering, metabolic pathw/ay en-

gineering, forced evolution, and adaptive mutations.

Why might natural genetic engineering be useful in modern

mrcrobial biotechnology?

What approaches can be used for the preservation of microor-

ganisms?

MICR00RGANISM GROulTH INCONTROLLED ENV:RONMENTS

For nrani industrial pr'r)cesscs. inicroorgani\nrs rrru:t ht gr,.,"i n

using specificallr designed rredia undcr careftillr contlollcilcondiiions. including te mpelxturc. rerlti()n. and nutrient ieed-in-e. The developnrent of appropriate culture nteCia and tlle

!f()\,, 1h 1)1 rliar()()r:-iini.llr\ Lmder rrlrltrrtriril etlntiltiotls llrd the

rLrbicct' r)1' lil\ \aaii!rnI3.'iolr'1,'1 ,,.'1'q',lrnt. rl i\ naaai\:rr\ to eilrjtr lailltilloi(lg\. Thi

ict-nt ltrntttrtation. i:'i'ri tn lr Ph-r:iolLrgical \cn\L' ln arrrllcr-ra!ti\rrtl,rl i]lq l)1rirk. r\ anrIi(r\t'tl iir lt nruiir nrot't gettenrl ,,rar ttt rel:ttiott irt

intlLr.trrrrl nri.ir,broloSi linil hi()teahnolrigr. As noiutl itt table -12.7.

the tarnr alilr h:irt sercral nrr-rrning\. rneluding thc nrass ctllttrre (linricrorrrguni:nr\ (r)r c\en plartt alrd anintal cell\|. Thc derclopnlc-tlt

of industrial ternte nlation: rL'qLlil c\ uPprr )prl]lc uulture llle dia alld

the large \cale screening of nricrooreanisrl'rs. Otien r enrs are neede d

to achieve optillrunr product vields. il{anv isolates are tested lbrtheii abilitr t() \\'nthc\ilr u lre\\ prodLlcl in the iicsirecl qr,lantrt\'. Fcu

are succe<.iirl

Medium Development

Tite itie.liuill .iiiJi,.r--ji,i,\ -r iill!T(,,rr-g.irli.tll i.,IrLrr:il ilc-cllil:.- ltCltll

influence thc :iolrLrrrre crnrpetitir enes5 oi a p&liculi,u proaa\s. Fre'qucnil\. lo,,,,ct-a..1 anrLlr nlntcrillls are uscd i\ sOLlrces 0i carbon.

一蓼

Fermentation:

「

_

Microorganism Growth in Controlled Environments 9■

429

3

4

6

A hrVord with Manythe Microbio t

An-r prtice ss involvrng the mass culture of microorganisnls either

aerobic or anaerohic

1n,1' blological process that occuis

Food spoilage

source Ra,Materl11 _____―一 ― ― 一 一 一

~~~一― 一 ―

Ca:-bon and energv \'{olassesWhey

GrainsAcr icultu[a] wasies (corncobs) '

Corn-sieeP liqutlr

So),bean meal

Stick liqgor (siaughterhouse producrs)

Arntnonia and atln'qniurt salts

l- itratesDistilier's soiubles

Crude preparatitlns oi plant and aninlal protiucts

Crude ino:qi11llc chcnicals

Chirlk or crurie cal blrtlale;

Fcrtii r zer-graile PhosPhates

Hightr alcohols

SiiiconesNaturill e \tal s

LlLrcl artd r tgetahie oils

ヽitrogen

irr the abscilcc of O.

fhc production of alcchollc bcvel ages

Use of an organic subsuare as the electrr:n doror and a'cePtoi

L:e of an organic -\ubstl ete as a reductanl and of the saine partiaill

degaded orgallic substrate as an oriilalt

Gr,r.tth clependent on substfate-level phosphorylation

,1い ｀`ll｀ t()P10(ltlCtio11 01に lCSil‐Cd lllctabOlitcヽ

Grovヽ√th Of MiCroorganlsttSin arl lndustrial Setting

l'he O:.()ileelltliliioll lrn(1 1'lLl\ rillt llltl\! be 'lll'li'iclltll illgh tir

hl, e (l- ill .:rccss \\ ilhill lhe cells so li'rrit ii i: not 1;nritirlg. Tili: ls

arsllccilill\ ltl.tc $ ht'll ll de llse iuic|ilhjlrl 'illturd 't gl'1111 111g' \\'hctt

l'ilarncntttrrs i'r-ingi and iictillolll\ cctc\ lila itlltLllcille ration cilrl ilt-

c\r-n lul'thcr llrliitcd bl l'illrrllentoLls sr\r\\th ttigure'12'-ll' Such

filarle ntotts gtou th fe stlils itl a viscr'ti: 111rL:tie rlle r'liLlill' kno\\ 11

as 11 non-Ne$ttlnian broth' \\ilich o1l'ei\ d\cll IrrllL'IL-\i\i'1ll!( ir)

sti rrittg llll(l ilerllli()ll.It is esscntial t() lisstlre that Lht':e pilr 'lclil 1'lr''1or' lrlt- llot litll

i,i,," ,11!('rohiltl sron th. 'f his i\ tlt"'l ''riticril tirrllrll scaleup'

there a sLlcce ssfill procedLlr- de reloptcl rtl li strr'ill shakc lllrsk is

nrodificr-l lor use in a iaiqe 1'erttletltet ()t.ie lltLtsi [lnder\tiil]d the

microell\'ilolltlletrt tl1'thc'cultLire lttld tllliitltaiti 'intilet ctlllilitiotl'

neartheindividualcclldi:spitcitlcr"lise:inth':tltlturcrirlttt-ttt'I1'a sttcccssful irailsititln cllll b' tllade it()lli 11 fr()'L-\s ()riglnitll\

Figure 42'4 Filanrentous (irosth During

Fe-rmentation. Filatl'itltlttLr' ltrngr rrntl sctinolll\ celc\

can change thsir glouih tornl tlttring tile course ola ier-

rnentalio;. The clcrelopr.nent ol pelleted grouth trr

fungi has nlajor L'tl'ects on ()\\'gen transfcr ancl energ\

,"qii,",l to a!itate the culture' (a t lnitial culturc (b) \l-

ter l8 houls sro\\ th'

Vitanlin.,c'Ir()n. trace siilts

B uiiers

Antifbani ltgents

fvtrj* Co*Ponents of GrowthUsed in lndustrial Ptglgsses

けChapter 42 lndustrial Microbiology and Biotechnology

Figure 42.5 lndustrial Stirred Fe rnienlers. ra ) Liu::r 1ci'

illarllal' u.e,l b) ii ])ll.rr'nlrLtIir.a] it,ntl.:l|ti 1irItltt nticr0l.rl] itrr,-ducliOrr Ul',rnliltioirt.. rltr I)eitriir rr1 il 1L-rntL.nlat Ltnit.'i-lri. Lrnit

cen l.c nln urirlci lL'r'!)hia ,)L ;ilt.tL'r'()LlL conclitiort:. rtnd itutiidnlittltltit0tl:.:arrrltlrrg..rllrl tr-nIrlt1.t{ion nt0,,it0r-in! eltrt l.c caIr'rcdoul Ll|(la| 11\al)t rt e\rrldil j()ns. Il i()\rn\{)r-s antl inl l -tl crr nli rl| [{ rt-r ilreatt ttrttridc riitl linrt inlolntltlion ()lt thc c()uf\e rtl lhc ltrnterrlli-tiolr. Speril-i. :ubstrllts. rrrelelrolic iltlcrrnediate:. alttl linal pr-r,d-

LlCt\ rlill b.' rlr'tcclctl

tle rt'loptd in rr l-i{j ntl I--rlcnilcrcr lllisk tL, rr l0( 1.()0() litcr r::ii,tor. thr'n thc t-rrof C:r i)1 :alilaUJr hli. t.r,r,lt I iill-]erl ,rul f ritIe rlr.

\1re lrrLrulinisrn\ Li:it ire qto,.,, n r:t crrllurc lrrhcs. shakr. 1'llisks.

Itncl :liilccl 1c't'ttttnlcr'' ()i otitaj illlr\\ LLl llLlta \\ \Lcnt\. Stirrctl lirrllL'iiL.l\ eiln [l]l!e iir 'rze llont.l LI J li1.'r: to liX).000 Iitr-rs rtr'

llu'rei. rlcpcndirrL on lr'rrductjr)11 trrlruiL'ntclrl\ {{igure -12.-it ,\tr1ri1;11 indu:trial \1nrdd i'arntcnliitl()n unit i. illLi:trutcil in f igLrrcil 5/,. \r,t,url., ilL:st 1]1. nterliiillt he:l.,rillu.-tl lirLtt tar'.i11()n. i)HJcl.iir.lnteni. .:ilI1t1i n.l. liltil ltr rtc,:.s rtl0nilirIttlg lltLt\i bc cet-r-i,:il

r jLlt LrnrlL'r ri-rolouslr clntrolletl eonrliltorii. \\ hert rrqLrired. ioaltL()ntrol lrgcnt.\ lIr.r\t he lrtided. r.\JleciJll\ rr ith lrigh Itroteirt ntr'dia.Cttrttptrters arc conlttonl\ u-(ed to ntonitor outltut( f ilrn probesthut deter ritine lliu r(rhiel bio1111155. lr-r e is ol critical ntetah()licprod,lcts. p.H. inpul antj crhaLrst gus contposition. and other pa,ranrelers. Environntenllil conditittns can Lrc changed or hcld constant over time. de penclin!. on the -goals f()r the particLtlar proces:r.

Frequ.-ntlv a e I il jatl e ()litp()ltent in ihc titctliunt. olicn the clir-boir slrrree. i< li.i,l..il a()niinu()Li\l\ iontinuous feed.-so thatiht liiiarLrrril:ilti:nt ii tll ::,,i i:-rr i. i\!tL'.\ .;-th.ii.tii lrr:illlrhle ltl anrli',r-rl iittlt l-hi. i. !:1iir!ri,:1 lr 1;11|, ,:[.i',] .,t'ir Iil.,r:r- iind Othl-rJilrhlrltldriilai. II a\1i.. Liriao\a i\ J,.r'a\anl .ti iht br-sinltirr:: ()f a

1iL'ilrcntrltron. it can be ratabLrliz.-il to r ie ld e ihlnol. rihich is lost

Culiure ornutrient aciditicn Dissolved oxygen prol

Coolingwaterout

Air in

lu: l rolrrtiic lrotlLrr't uitrl r-etiLrces thc f ilril lielil. Thii clln ()cc1

i'\ rn rrndei ltc'r'oirit c()nriition-\.Ilesiclc: llte traciitionui stirr-erl aer't)hic ()r unal-r()bic f'enrenle

othcl lrpprolrchcs cirn Lr.' uscrl to gr(i\\ rrricli,rrganisnts. These a

ternlti.,cs. illristrrrtetl rn figure 42.6. includc irtt-tube i.nlenlertligirlr' 11.(rrr). uliiih clintinrite thc neecl li,r:tirrcrs that can L:

loLtled h,r'f illutcntoLts 1Ingi. Also arailahlc. js:rrlrr-1-state ferrnertrrtjon Iirgulc -+1.(r/r. irr * hich li plirticulatc \ubstrate rs keltt ntoirto nlrrntliin;t tlrin sLrrl'ltce riater i'ilnt uhcrc nticrtibes ciln gr()1

rnrl orvgen is aiaillrblc. In rarious l1'pes oli'i\ed, (l'icure.+2.(rc

and t-luidrzed-bed reactors (figure 42.6d)- the micrtrurulnj'nts lrassirciated u.ith ine'r-t surfaces as bittfilrns (see pp. 6(1.?--l). an

nredlurrj f-lou s past the fixed or suspended particles.Dialysis culture units also can be used (figure 42.6c.t. Thes

units allor.r toric wliste metabolites or end products to ditfusau av fl'om the microbial culture and permit neu,substrates to cliffuse thrtru-rrh the nrenrbrane tou,ard the culture. Continuous cultLrrc tr-chniqLres using chemostltts (figure.l2.6l) can ntarkcdlinrpror e ce 11 ouiprits liltri rttc\ ()1'srrhitrate uSt br-Cl1u-sc ntict-r)\)r!iini.ln\ can he ntlintained in a continut)i,ls iogtrithnlie phase

1-{ou cr e r. continLi(lrri ntaintenancc ol an ot.gani\nl in en litir,gro\\th phase is undesirable in rnanv industrial processes.

42.

ｅ

一鏑‐ｉｎｅ

972

pH probe

二

・ 42.11

{a) Lift-tube lermenter

Density difference of

gas bubbles entrained

in medium results in

flurd circulation

Microorganism Growth in Controlled Environments 973

(b) Solid-state fermentation

Growth of culturewithout Presence of

added free ',ater

(c) Fixed-bed reactor

l,4icroorganisms on surlaces

of suPPod material:

flow can be uP or down

(d) FluiCized-bed reactor

lv'1!ciocrganisn-ls cl sur{aces

o{ particles susPenCecl

in liquid or Eas strean '-upwarC:iovr

FlovJin+:

―Ar n

FloヽV Out

F orv out

SuspendedsupPorl Padicles

FoW n 一

(e) DialYsis culture unit

Waste Products ilfluseaway from ihe culture'

Subslrate may ciifiuse

through membran€ to

the cuiture

(0 Continuous culture unit (Chemostat)

Medium in and excess

mediLrm tc v/aste \Yittr

vJasted celis

Figure 42.6 -{ltcrnate \lethods ftrr }lass

(.JIture. ln atlclitiorr to stirrctl fr:rnlcniers' othet

lrr(llt',r1. . .ttt irc tt'."1 i(r L rlllrllt' llli' i""t j'tlll'ltlt ill

urr-ltt:tnril ilr()cas\ci' lti Inliilr 'ases

tilLlse altel'nale

lrnlll olichc' \\ il1 ha\ c 1(r\\ er ()lleratlllg costs alld. car

1,,.,.,.t. .irc'illiztJ !l ()\\ th'()ilditi(rll\ tlcecletl i'tlt

Ir rrriLrcl .\ 1lt1-la\i\

1ビ lalion、 hip to lhCヽ 'ゝ1lthCsiヽ ()「 cCll 11latcrial、 alld l10r,1lal groヽ 'th

lヽoヽ t antibioticヽ anci thC lllゝ COtO｀ ins fall intO this catC80ry

t. Uo* is the cost of media reduced during industrial operations?

;;;r;t iire effect of changing balances in.nutrients such as

tr4edium rn-*€

\ Iicr-ohi;i1 Pltiduct\ tlltcn arc cla::ifit-c1 as prilllllr\ and :ec'

ondar\ metabr)lite s. As si.torvn in figure t12.7. prinrar] metabo-

lites consist of cotripouncls reirtcd to the s\nthesi-i of migrobial

celis in thc grou'lh phase Thel inciutle ainirlo.ilcids' nucieotides'

and f.rrnentation entl orc"it'cts stlch 3s ethlirlol rrnd organic acids'

In adLiition. industriailr us'1'Ll1 L-nz\ 1l)!'\' e ithcr rrsttlciatecl u ilh

1|1g 11i;r1rl1ial scil: trr i\t'c1l/)111r" \'1t'll '11'- \\ilthc\iz'i fr iltr-

cr(rttt !:ttli:nlr,1url ilS !rL'ri tfi'

Secondarl metab()lites u\Li;illi ii!''tirlitllJi' jLititl: thr i-t-

rir-rrl ot nutriL-ttt litl'titatlon t)J \\ lirie pttlrii'tcl 'tiii:lll''illtitltl that iLrl-

lo$s the acti\c sro\\th phrtc The:e ctrlllplrllnds hare no ditect

minerals′ 910ヽγth factors′ and the SOtl「 CeS Of carbon′ nitrOgen′

and Phosohorus2\.^VhatarenCn-NewtonianbrothsandWhyaretheseimporlant

in fermentatrons?

3'DiscussSCaleupandtheobjeciiveofthescaleupprocess'

「

chapter 42 1ndustrialヽ 4i(robiology and BiOteChnology

5. Besides the aerated, siirred fermenter, what other alternatives

areavaiiabieforthemasscultureofmicroorganismsinindus.trial processes? What is ihe principle by which a ciialysis culture

What parameters can be monitored in a modern' large-scale

industrial fermentation?

system iunctions?

MAJOR PRODUσ S OFINDUSTR:ALMiCR08:OLOGY

Time

pre 42.7 Prinrarr anrl St'c<tridarl \letahrllites' I)e-

r,1ii,g ,,n tiic llalricLrilrr-otrllllll\llr' ihc tlt:ii'ril llrorlltet tlllti bc

,ra.al .1r.,ring or a1'ter o""tth Pi 'iirlrr tllttltboltlts lllLl 1(rlllle(i

-ing thc ,,ati, e gro'n'th ph:l-'c' u ile tclts st'c(rll(l'lr) lile 1'tbLrlite s

forrttetl atier grou th is cirllllllctctl'

Iniiustrilrl nrlctc,L.iolcigr hlts Ptt'rr idt'd prodLlel's that haYe in pro-

1'oLrrtCli' cilirngcd iir-rr li" es :rnri iiie -tpans The y include industrial

lnd ,.rglictrlrLrrlil prrtduclt' 1'ood lrdditir es' iliedical products lor

lrun,,,l unci rinjllii 1-rtalth. rtnd i-'ioir-rels itahle'12'9i Particularly"

in rhe iasl 1e',i r erirs- llonillliibi()ric !olllp\)rtlld-\ rtsed in metiicine

ancl healtit hure tliltrlc tilrijLit c,,tlitihLltiorls ttr the itnprrlved u'ell-

beinc o1'eililllal lrll,l hilitlrln polluf iiitlns' Onlr l-tllijor products in

r-i-re h arite!(rl-\ u ill lle tli'' ti'<cti ilr-rC'

Antibiotics

\lanr lrttiltrrrlla .lla 1)t'rtltltr'ti hi tllltl(r{rl!Jllislll\' preclonli-

ii1i1-,i]r lri iiiiiil(rill\ltl.ls :ll tht geitLts !/r1'r711r17;-'tt'' lrld b)'fila-

,,,a,,,,,u.'fLItlgt i.ii'. tirhlr' -i-i 'l' irl "l

' ; ehlllter' thc irnthesis of

seretlll i)1 tlrt'Ill(l\1 lllll'1i)llrrlt ltrltibitttl-: rrill be tiiscusscd to il-

ndustrial I'roductsEthanol (froin gluco-se i

Ethanol r:from l:lctose )

Acetone and butanol2.3-butanediLrlEnzl'mes

rgricultural ProducLsGibberellin-s

'ood AdditiresAmino acids (t.g . 1r sine t

Or-canic acids (citric aci'ilNucleotides\4taminsPolysaccharides

vledical ProductsAntibiot'icsAlkaloidsSteroid rran.'rfonnations

lnsulin. hunran grou th htlnlone' som3tosi'iir

BiofuelsH1'drogenN'lethaneE:thanol

-Sur'aftrrltriltr at': t {'r?1 1Jl(?('

K Itttt c rontt r t s .fra gilis

Clo st rid t Ltrrt ttce I ctbutt I i ttrtrt

Ent e t ol.tatt e r, St rrai ia

.{,t1rcrgi11iir, Bttt.iIIus l\1tt{ ir' 7 tt'-lndIrn1Lt

Ciltbtn'![o.fujLkunt

C o n ne btttt t' ri tt n 3l ul arn i t t tttAspergiLlus nlgr:t

C or : ne bat'le ri tt ttt 3l ut tt rtt i c tttrt

,\ :hb: tt. E re nu nlrc r'i ttnt B I ttkt sl t' tt

Xtutllrcntotns

Peni t' ill ium. St rep lontt.<:e s' Bac i I lt t's

ClavicePs PuryureuRlrr:,,r,u' 1'111"'l'a' 17r

Est Jier ichia t't',Ii \titcJruronu "s "/1'l lsl(l''

.;-., ith'r:,:' tr\A 'e' l-n 1 '':i

Phuir:r nlhctii miir,'itr:slisl:',\1 e r I i tt nt t b ti c ! L t i u nt

Zt'momon as. T he rnn onae ro bacl e t

and oth:i-s1「ltCl■ CrOnヽ

:' Prirnary- metabolite

な）

formation

Growlh ,/

、、

ヽ ｀有

; .." Secondary- metabolite

formation

Major Products of lndustrial Microbiology4213

（ＴＯｒｘ

ｏヽ〓くの）Ｃ〓〓ＯＦ一ｏ０

ぃ０ア』ｏＦＦＥｏ

０“０」０ゝ〓００」ＯＯ

．でｏ〓ヽご）のりｏＦ一〇一ｍ

o ?o 4A 60 80 100 12n 140

Fermentation time (hours)

Figure 42.8 Penicillin Ferlrtentation Iltrolres Ilrtcisc(ltlntrol of Nutrients. 'l-hc s)'rltllcsis o1'Perricillin hrgills n lt"rl

rrittt,-le tt 1't'titlt ltlttttlilt-tia becoltler lirnit;ng' '\1-tcr ltltltt ot'tht- lltc

t6s.. rlr sipl 1r cutahplizcd ilislrer'hlrt iclc)llri: fictlt dcgllded. r1ir

cr)sLt (it rlrpidll Ltsctl tlltlllrlslicchliritle) i: liddctl :lloll-l rr lth 'i lorr

icr el ,.rl- nitritge n_ Th js srintLtlate s nL[.inrunt tt litlsl'trrtrllititllr r)lthr'

cai-ltLrlt \otlrccs ttl llclliciliirl. Illt scllit t'alt()l i\ Irl tst'lrlttl tIitl-

the crtlt\etttii)n fec()illlllrrlldctl br t lle'\SNl"iilat l5' 1 11li111[1'r. t"t'i

the aris shoitld be IlrLrltiplii:d br'0.10 to olrtliirl tilc trttr'ralLte '

lustl‐ titc thC Clitical role of incdiuinn forlllultlti()n and cnvil‐olllncn―

tal contl‐ ol in thc productio13 of thcsc inlpoltant colllpounds 蠣ヽ

tll),i ti、 1 ,t、 、lit t lt■ 1ヽビ: 「` |

Penicillin

恥ぶ[1鮮議I勲熱、11#:Ⅱf鷺

i研硝‖篤胤烹博 諜∬だ11ゴ賓I葛Iぶ

‖窯∬)点‖I鷲∬fttIIT∬‖鷲Fζ‖ボヽ性R満樵mtibiolic icヽ ctt inぬ c culm「c hcginよ iヽn劇で■、ctngurC 42.9,un―dcl ct,11こ liti(、 11ヽ いic● 111「011Cd 1litlo=Clユ li1l111[ttiol〕

Anlino Acids

へ1ュ 〕i1lo[lC iti｀ tヽlch ti、 1卜 、111ビ [IR(1ピ 1ヒ ltユ 11lic tlご iti ttせ ょヽCd in tllc lkヽ いに1

ilndtl、 tlヽ aヽ 1l tlt■ tl● I〕 tilヽ upplclllcnts in blctld Prて tヽltiCiS tlind tl、 1lti、 or―

cnhancin=ご 、)11lPい und、 、uch t卜 1110n()ヽ odiunlピ lutanlatc t｀ lSGI

Figure 42.9 Streptonrlcin Production br

5tr-tpt,tmrctsgri-serls. L)tIlci"''- "i jiii''i':' lelii: 1o

nriL\iillunl antihiotic I icids.

０●一●

，

〓

。

３

Ｅ

ＯせヽっＥ

】

Ｅ

５〓ｏＯか

，一つＥＯ

（，

こヽゅ〓し

ｏＯｏ●ョ一ぃ

いＯ

ＣＯ〓●Ｌ”‘ｏ●●●０

奮Ｆ一ヽい、）Ｃｏ■ｏし″Ｃ●ＯＣｏ●

●一０、Ｅ

Ｏ″αｏ」ヤ∽

975

― ―

Chapter 42 lndustrial Microbiology and Biotechnology

ｔｅ

　

　

　

　

　

　

　

　

ａｔ

ｔａ

ヽ

１

１

１

‐

―

ァ

‐ｔｒ

＼、、、、　　　　　　軒『一′

a)

le hv Coi-r'rebntterirtt'rt qlulamicum. \ 1a.jor cer.hon tlrtws arc notcd by bold arrows(a)Growth with usc ofthc gi)()、 卜late bypass to

shitjccl holtj r|.ro$s) 'l he dlisiterl line-s ind;cate rL'lcti()ns ti-litt lre bcrllg used to a lesset e\tenl.

42.14

ｓｅ　　　　　ｏｓ

∞―十

ｐｈＧ‐ｕ　　　／／／／／一

ａｔ

　

　

　

　

　

　

　

　

　

　

　

Ａ

ｐｈ

ｌ

ｌ

十

ｃ

T百 ose pho

|CO

/ > Acetyl-coAC

CO

封r

)γ

」:te lileAcetyl-CoA

CHOC00~

Glyoxyl21s,,

′Oxa osucc nate

r

●Kao― ″・島

91utarate ‐

INH+

T

G utalnate ti))

Oxaiosuccinale4.

,,-n",o @*& UU.

g NH.**w

Glutamate

＼ヽ

Fumarate

I\\lr

Succinate

\ \Suce 1nYl-CcA

I

lsocitrate lyase

cis-Aconitate

\I

I- - - lsocitrate

&6sF

.!Irn() :iai11 pt'orittctton is t1'picall-r' cal-ric(i ()tlt b\ lllcllrl\ { )1

latorr ntutatrts. u lricit itavc lt redLlct'd ahilitl to liitlit svn-

. rr1 liir r'ttLi pir)rlLlal .f

h.' ltrlrnlal illicrool-lliillisrl ltr oitls tl!e r

l.ti(rn ()1'l-ri,rlhe lrillrl inlr:rnreriiltes b,r'the carefirl lcgulllli(lllllulur rlcirih,rli-ittl l'roduclion Lrf giutarllic ae id and scr eral

rllrnr)i1ai1i\i\Il()\\ al1l IiaLl outLlsrllgllltltxntsof ('rlr-t'llrl1.;ttt-

t: <lirtLrnitLirur tlllt leck. ilt htirc onl\'ll linlited ahilitr tcr

'cs. ti're lL \ crule IIttct-ltlediiite u ketog]Llla|Jle (.\ce dpPettr) t(-r \Ll.r'ln\ l-(lo-.\ as sh,.rwt.i it.i figure ;12.10. A ctlntrolled

riotin lerel uird the eddition oftattY acid derivatives results

:reasctl menlbrane permeabilit) and excretion of high con

:itions oi'siutamic acid. The impaired bacteria use the gl1'-

iie path$a)' (see set:lion 10.6) to meet their needs for

tial trrochen'rical interrnediate s. especially during the grou'th

'..A1ter gronth hecotle s lirnited because olchan-ged nutrient

:hilitr. rin lllnosl cott.tPletc nlolar conversiot-t (or 8I .79

tl a()n\er\l(ri) oi i\oaiirait t(l !luianlate occLlrs.

\ \iila. ilir a\scntiiil :lllllll(r llCltl usr-d 'ra) \ullPlerllent alefeal-\

rrc.1d\. '\ :is oriSinall\ ilrodu.e d in 3 t\\ o-step nlicrobial

PrOCCSS Tll:、 ha、 bccn rcplaccd b)を 1、 i llぎ Ic― tヽCp it「 IRlcnttltion in

、、hich t1lc l)actcritllll(｀ て,′l、′7こ′わで

`(rピ′′ι″77 .、 ル

`た

′′/111′ ″だ い1● ckcd iln

thc s、 1lthc、 i、 ol:honlo、 crillc ttCCtlllltlltltc、 1)ヽ lI〕 c ()ヽ cl■1=llitCI

Ctll]bC prOdtlccd iin t1 3(la、:fcrlllCnttltioll

Organi〔 Acids

(Drganic acid pl‐ odtiction by 11licro● Igtini、 l〕 1、 1、 111lPい lttillt ill i11-

dtlstrial 1lnicrobi(Dlogy and iHustratc、 thC cllicct、 ぃli ti tiCC I:3Cttll lc、 ―

cis and balanccs oln organic acid、 )ntlncsi、 and c、 crctiぃ Ii Citlic

acctic,lactic、 fuinaric,alld gluconic aci(ls arc nl轟 or prOducts

(table 42.10)Until nlicrobial processcsヽ crC dC｀ cloPcd tllC ina―

jor sourcc ofcitric acid was citrus tuit fronl ltalゝ 'Toda).most cit

ric acid is produccd by microorganisnls:70 r iヽ ヒl、 cd in thc food

and bcveragc indust]、 201を in Pharnlacculical、 and thc balancc

in othcr i1ldustrial applications

Thc csscncc of citlic acid fcrnlcnttitioi〕 i11、 |,1ヽ じ、li:lliti:1ビ thc

a11lounts of tracc nlctals such[1ヽ 111■ 11=tlll C｀ C t11こ l ir● 1l tt,、 tい

「 1ヽ―

/'ど′メ′/′″

'″′gど ′groヽ 'th at a sPccilliC lっ olllt ill thc icr,1lc1ltation l‐ hc

ゴヽ

'( > Aceiyl-CoA

CO:ツЮacdae / 6tde

Major Pr-oducts of lndustrial Microbiology 9774215

fVtrpt Orgrnic Acids Produced by Microbial Processes

Fermentation Conditions

Acetic iicid

Citric r.rd

Fumaric acid

Giuconrc acid

Itaconie lciti

Ko-jlc acid

Lactic acid

ism Used

Ac e t ob(1( 1 e r s'ith elhanol solutions

,4 sper.qi,'ltr.r rt i-ger i r: l.toias-ses brtsed

nredium

Rhi:opLts ttigrir:ans in sugar based

mcdiuin

Aspe r';illus /i!'gdr in glucose-inintrai

salts mediunt

..\;ptrqillu: lt'rrctts in mol15se\ salts

me d;un-i

,4 s pt rg i I I u s .fl tt rtrs oir'-rre incarbohvdr:rte tntlrgrnic N mediillll

ilomolirmentati !e a,i.1r,(l.tllll t

,it'l brur: t'kii

tatiYe Uses

Yr'ide variet-v oi' iood Llses

Pharnaceulictlis. as a lirod itdditir t'

Resin man,:lacture. talli'ling.:'rnrl sizing

A carrier tbr calcium and siidiurn

Esters can be Prtll''rterizedto make plastlcs

The manufacture ol fungicidesand insecticides u'i.ien crltllplexedu ilh melels

.\s a . rn-ier tor calciunl and

as an acidifier

Single-step ttxidation. *ith l5%solutions produced: 95-99% yields

I{igh carbohl'dr:lie concentrations and

cuntroiied lin.iitation of tr:ice metals:

60 80% vields

Strongll aerobic tcrmentation:carbon-nitrogen ratio is criticll: zinc

shoujd be limited: 60% f ields

Uses a-eitation or stirred fer:llenters:

9-i7r yields

I-lighli aerobic medium. below pl{ 2'2

35')i. vieldslron must be carefully controlled to

arojd rerction tirh kojic acid atier

i ernrcntationPurif iti mediunl used rrl tliilitate

eltracl iL)ll

Irrr-dirinr rtitctl i: treltlr'd utth totl d\uh3ll!e l.'tiil\ i!) 'll'tllc

l()\"

;jn(1 L()ltll!)llarl roitcr:lltflllir)11S ()i'lL\illlilllle tllCtlil'. illl!l l\ aillli'ri()ut in iiet.(lbia .tit rcd ielnt..nlL.t \. clerlL-rltilr. ili gtl \'l!iil L()ll! rrll

Ll.rii.'r.,ilr : r'r' rr'r'1..1.i,,'1'1,r11f i"ti i'i: i:' \''riri

Ierlct tllL i]lhii-tti,rrr oi eitrii li.id lll)tiLlctlon 1), il.t)il lrb.rt'().1

irprll. I hc stlrLt\\ Lrl thts lcrllltlltiitioll tie Ilt'nli: otl tht ltgtllltti()n

rrnci lirncti0ning ()1 titc ulr c()l\tic pilih\\ l\ lltlti 1ll.- Itie ltl ilirr) lit'

ilai(l !\cle 1ir.r' \{'(lit)tt !i.+)..Al'tcl tlle lLciirL'gl()\\1ll l)hli\'' \\lltll

1113 :11ir:1t.rrtc ler tl i' lirril. ciLI'ate \\ nthlst' ilLti\ it\ iilclclt\L'\ lirlrl

tltr- lriti" i11L'. {)t l.i(i)llliil\c illlLl i:0cilretc tlchrdro!e lllLsc dcrr':lL:c.-[]11'

1-sittlts irr iitiic ilaiLl ilucLilllLlllititln arlrl c\'1r-lioi'l hr the

sllL s.t'l lt,'' I ' :-.llri'll'.In !,)rtti.r:ii ir,rtl. iilt' llirrdLttlltrtl ()1'-Jlll'i)llic ilci(l i11\ rrl" l: lt 'itr

glc tlllr:rtrhilti cilzr lllr'. glLlcot3 o\idilsc' ilrLrnti tn l t/ri r'(tliii'

niqt'i: .1. rilqri l: !l()\1.11 Ltniltr (llltilltlllll coll(li1i(rll\ itl r '"111-.iealt jt(lu(rr ntctlir.till (rrtlirth bectltlles lirllrtctl br tlitl-i'!elt lintl

ihe rt-stiilg aall\ illlll\lorlll tiic lclllaillinS glLteose t'r !lLtcrrtlii 'tciil

rrl lt iril-!lt-slal) lr'11a11(r11. (ilLrcrltlic eciLl i: irsctl ii\;l (rilrri'l l'01 clrl-

aillill rlllrl i1(ril iiil(1 t( il Li)lll|()llr-llt 1r1 (ict'lgell1s'

Specialty Compounds for Usein Medicine and Health

ln atldition t() the trlllk pl odLlct\ that hare ilcrn prritluced tircr

the lasr -l()ro '1() \'cars. such a\ antibiotics' arllino acid\' and o1'-

ganic acids. illicroofganisljls alre u:ied ior the orodr'tctioll cli'

nonantibiotic specialt)' conlpounds. These include scx hor-

nlrlnL's. linlittllllor sgents. ir,noLthores. antl spcciltl '(llllp()ulld\

ihlt lnllr.icnce bactciia. tunri. atlrrehee' in:!-ct) iind fllllts,table-t2.l1r lrr lrllLlt:c). ll i\ llL-!c'\t':l:' t' ir"dilil rrittl Iitrr\il'ihe r,t-,,ttLicl. itirdci iaicl'rilli .irn1ri,11ct1 cirildlli'rir' l1r:i\\r'lij illlll

'Lht:t;r-lr-rilia]lr iillPt'rie;ll a(lll'lll(rtiIlLjr fL'llth illc'-rrll'!; illJT ril 'l

rrihlt. r1ltr l'\ i r "l'lJlli'rll.

1 What critical Iimiting factors are used 1n the penicillin and

strePtorni'ti n fermentations?

What are reguiatory mut2nts and how were they used to in-

crease the production of glutarnic acidby Clrynebacteilurn?

What is the principal ltmitation ci'eated io stimulate citric acid

accumuiatron by Aspergillus nrger?

4 Give some important specialty compounds that are produced

by the use of microorganisms

Biopolyiners

Biopol\'mers ltt'e tllicroi-rilill-r' llrodr'rced p()1\ illeri' prilllaril\i

1.,,lrsli.:ch,iri,l.s. il\ccl 1ri gttrdii) the ilo',r chtrlctcristics oi iiil-irirlr utt.l t() scl\c Lls ccllill! llgcilts. Thc-se llfe eniplr'r\'ed tn ll-tatlv

.rrcls o1 thc phar tllltcctlticll ilild lootl jndLrstrie: .' I

.\1lelr\lli'ioiallpoivsacchltt-ldesarcuscdirsstlihilizers'forilrc .ii:pr'r:totl ()i lrilrliltllllte-s- lrs f ilrrl-iorllling agenl\' or 10 pro-

ir(rlc ',i lte t t e tetltiilll ill I at iou: products. Ptrlr''lccharicle s he11l

ntiiintuin thc te \turc ol nlitn\ 1'rozcn lixlds. stlch ils iCe Creitlu. that

1i1.1 511f-iect to tlrasiic telllperatLlre challges' These polr:saccha|rdes

musl rraintairl their properties urtcler the pH conditierns in the par-

ricLrlar lbod and be compatible \\'ith other polysaccharides' The)'

sl-rould not losc their phy'sical characteristics if heated'

Biopolvmers also include (l) dextrans' which are used as

blotrci .jrpalrders atltl ahsol'bents: (2) Entinia polysaccharides

rhat lr'. us.d in plinis: anci (l) polvesters' derived fronl

F)\(,tLlttni()nti.r rrlr'r,r'rr'au1.r, nhich are a feedstock for specialtl

p1;1s11;r. L'tllu1L,.t'tlrtci(r1-illril\. plodLlcdd bt'trt '1t'ctoboc'tcr

.illin. rr. it\..1 r\ ll 1'r,,id thicke rle r. Polvsaccharides such as scle-

i{r!lLtcllt ale trsed b] rhe oii intlirstrr as drilling nlud additivcs'

t,

97e Chapter 42 lndustrial Microbiology and Biotechnology 42.16

Nonantibiotic Specialty Compounds Produced by Microorganisms

Pol),ethers

AvermectinsStari ns

Enzyme tnhibitors

BioherhicidesI;nnr u nos uppressants

Anabolic asents

UterocontraclantsAntitunrdr agents

S I re pt o mr c e.s c i ru ru t tn n e tt.s i s

S. losalien.sis.\. albus

S. averntitili.sAspereillu: krreusPet, ; c1 1 1 1,r,, c it rin unt I

aclinomycete'

S. clavaligeru.sAc'linoplanes sp.

S. hrgroscopit.u.s

To Irp oc I adi unt i rtJ ! a t tun

.5. tsuknbaensi.;

S. ltt e ru.; t r4t i t' L t.s'

Gibberclla :roeClaviceps puqtureoS. pertt:et ic u t suhsp. rrir:.irrri,5. lteucetiL.r.:.5'. i rzr'.tpilo-trr.s

N{onen si n

Lasaioc id

Salinom,vcin

LoYastatin

PraVxstaii.

Clavulanic acid

Acerho\i:

B iaiaphos

Cy,closporin AFK-506Rapaml'cirZearaienone

ErgLrt alkaioitl:Dororubi c i n

Dru noruh i ei n

\llrrrJl\arll

flleornr cin

Coccidiostat. rumenal grorvth prornoterCoccidiostat. runinal grori,th pit)n.toter

Coccidiostat. runtinal grorvth prornoterHelrninths and ailhropodsChoicsterol -lorverin-g lgen t

Cholesterol iou,erine agent

Penicillina.se inhibitor,Intestinal giucosidase inhiLriior klccre:ses h,r peigly cernra and

irislvccride s]'nthcsi-(l

Organ tr:rnsplanrs

Orean transp)ant:Organ transpllrnts

Farnr anintal medicaii0rInduction of laborL anaar treeimcntC:ncer treatilenl( itnaei trcrtlntnlL ancr'r lr i'lttntcnt

Compound Tlpe Source Specific Product Process/Organism Affected

"Compac(in. produced by Peniciilium citrinurz, is ch.nged to prr,.asElin bv iln aclinomvcete biocon!crsion

Based on: A. L. Denain 2000. Microbiai biotechnology. Tibtech 18:26-11: A. L. Demaii 20{X) Phrnir.eltri:it!\ a.l!e scrA- L Dernain. I999. Seconciary metaboiism in bacleria: Antibiotic pathways relnlation and Iunction in Bi.,l,r(r t,! th; pni:at

,nd!^' meebojites 01 microor,ganirms_ .1/r/r. Milrr,rr(rrl..r. J \\'- Len-q.ler G Drcrs_ and Il a;. Schleeel.

I Butahntl. i):155 .{'61: c Lancinredibrs 617-51 Nr\r Yorl: Jhicnr

-l\antlltn i-r()lvntel\ eitl]liltce ()il feco\r.f\, br, trtrl.;, 1\ tnLt \\.ltL,rf'lr)odin-e and the displaceruent ol'oii. 'l'his usc of xaltthun sLul.lll'otlttced b)'Xurttltontotld\ (LuttlL\//t\. tcptc\L-itts ll iarsrt ]t(rtant ul nrarkct 1or litis ntrcr ob jal i)r()duc1.'l'hc cr cl()d.'\irin\ har,e lr uniqLlc \truulLliLt. it\ sho\\ n illfigure :12.1 I 1or cr c],clodextt ip. Ther' are cr clic gliSrr:rictri:r,ritlcs u ht)sc sLr!als iire .ioinr(i br o I .J linkagrs. C-'t clotlcrtiinrcrin bc use(l litl ti uirie \ariet\ ()f Jrul'lloses be.caLi:c thcrc err.ircll ntolccriles hind u'ith suhstances and tilodil'\, tlre ir pitr sicllIlr()pcrties. Iror crantple. cvclode\trilt\ rr jll incrcase tltc :olLrhil-itr ol'phlrrrrurce Ltticals. rlraluce the ir bittentc:is. unal nllisk ( ]tcllrlciil odors. CvclLtdcrtrins als() can ire usetl ir_s sL-lecti\ g 1lr1r1lrg111r

t() r-enro\c cltolcstelol lritrit eggs anri bultcr. t, r []tllcrt \]l IL.\ll orrr oridal ir)n. ()i' us stat ionart' pilliscs in gas chrontitto!i ltltlt,\

Biosu rf acta nts

Biosurlictants are especiallr,. itnportant lbr envir.ot'rr-r-rental allpli-cations where biodegradabiiity is a ma.jor requiremenr. Ther, areused tbr entulsitication, increasing detergencl,. \\,etting and phasedispersion, as well as fbr solubilization. These properties rre c.-pecially irnportant in biorernediation. oil spill dispersion. and .r,hanced oil recoven, (EOR). The ntost widelv used nticrobiallrproduced biosurthctants are gl\ colipids. Thesc cornl(runds h t\ cdistinct hr drophilic and hr.drophobic rcgions. and rirc c\..lldltidispr-rsine agenls.

-l-he-r, have bcen use d \\'ith the l rrr.,r: lrrlajr.-

and other oil spills.

cylpS\-o- /r-v \

s1 PA."q:l^{/) "' i t*Sia*o,bo+ \\'

K,on Ho-,l'o..\ ryo9,o2) )o+_ t/ "t -g o-" 5to) | {[r---,-r- oJ o F

o',AO.HJ

ir-Cyclodexlr r-

Figure 42.11 .{ f'r clodertrin. The \irlrcturc rrl ,,-r'., clodcrlrin.llrtrduct-d ltx I'ltt'prtt,tlrtt(k)j)uLr(,1. Thcrc als6 tue [] 1;,i -v iirrlts. 61'

sinrilar \tniclurc bLit Ialger: these cor.npounds ha\e iltan\ applica-tions in ntedicine ancl industrr,.

Bioconversion Processes

Pr i, rrtr.rir', rl \ ii )il \. :tlrtr k n, r,,., it li. nlicrobixl t ransti rrntations r,r bi<t-transfirrntations..rl-1.;Ilin,,r ih.rn-ac. 111 1111ri3;ulg...r,Jtr ii\ lhr- in-

-\iitron ili .t i:rdiir:.i i trr kr,to liiniti0n ()t tllf \iitLtr.iltl]t)ide\itturaliOn,11'a ci'rrtpler crclrc stnrcture. that are can-ied out br nongro,,ring

L979Microbial Growth in Complex Natural Environments

4217

?'.C:O

Ho- ^. I -l-,i I-l'\J_iMajor ProCuct

Figure 42-12 Biotransformation to \lodifl'a Steroid' H1

ilrox-v-larion of pr,lgeslcrone itl ihc llrr position b1' Rhr:r;7;lr'r rli-

gri,',,,,.r. -fhe str:r,rid i5 il15-'t-'lred in accttlne belor-e ldilititln 1o tlle

ftr.-r,,\\ Ir itlllf :il . ttltLtlr.

n-iicroot-garti:rlr\.'l-llL' ttlicttlt'lgal.iistns iilLls lict li: tlitlcatallsLs'

BiitCorrr e r:tons Itirr e ll-lltllr atlr .intli!e \ i)\ L-l ChellliCill prOCttittres'

A nltjor adr rLrtllige is sterctle ltr:tll jcai' the hirritrticljlr ltclir e tirrrtl

o1' I prodr.ict is rltlttlc. [:llu t'ttlcs lt]str cltrrr otll \ ci\ :i)ccillc rtiic-

tionr utrricr nttlt1 rrrtlritlirll\. jlltl l.it'l't t'.ilut tll'illtthlt tlltllCcttlCs

clrn ht- tftrn:1,1I'n.tcii. l- ni.t1]Lil:il hlrtene. llatill()lt]\aL'tL'\. \L'llsls.

;rntl ntoltl:her..hCtIir:tJin\.1ilr)Li\lr1|q,rl1 -i'l()ll\.'lheCtlzrtltesre spttnsrbJ.. l0i. thc:c !.(r11\ L,f\1()n\ .etl hc liitlltct'llrllllr 0l' t\tflice i

Iuiar. cells 1'1111 hL'1tlrrtli.iteuL jn Lr:ttuil iil !()lltillur,Lis cLtltui.l allrl illcil

dried lirr dire,'t ust-. rrl lllc\ .irrl bc pre pltrc'd ilt tttole sllceif ic uavs

1,, \.,rf\ ',rt1 ,lr'.tr,',1 r'.,'. ,',\s1'j"r''.\ 1)'pialll t iiliritl', 1-l:i(i11 is tht hrdrorr laitilll tr1' a stctorrl

rligure 12.D.l ln this trrtrrpll' thc s arct t"t6lgLrle ste roid is

dissolyctl lp aCL-t(r1c erlii ti'i.'ll ldded tg tl-ie rcactiOtt systell thal

cotttltjns thc llrcurtlrl n rlliclrlbil] cclls' -lhc cotlrse o1'the rrrodi-

iicetiort is tttirrtittlrtri. rlntl tl'tc f inll ;rrotitrcl is e rtraclcil lronl ihe

tllr'Lllllrll llrJ 1',tt tl tr'i.BiOtranqf'onltat ions Cllt rie 11 oLrl br i'lCr' L-ll1)'llie S (lr lntilct ll()n

g11ru tttt uclls tio hltr.' ljnlitatiotls' I{clie liotl' thltl occitr in tlre lrb-

rc,tc" ,,1 uirjrr' ittel:il.olisitl .'r ithtrLtt leclLicin!' llor\e l r)l :\TP

hcing ar arlablr- a()lltllltl3ll\ - ilrt pt itnal'il\r cxcl q()lllt tcitL tir lll\

(sr'(' .t('( 1lr)i r ,t -j i. l1 {l-P ol rdrlLlct:lllts rile letltlired' sn cnerg\

':ource \Llch 3 gl111 ose ]lltl\l ilt'sirpplied urlcleI carclirlll'eiln

tr olletl ttotttr()u th arrlltlili()ils

'l Discuss the maior uses for biopolymers and biosurJactants'

2 What are cyclodextrins anC why are they important additives?

3 What are bioconversions or biotransformations? Describe the

changes in molecules that result from these processes

MICROB:AL GROWrHIN COMPLEXNATURAL ENVIRONMENTS

\,licrr,hial ntzlnaltrntcnt ul\(l ci]lt be carried ()ut in colllple x natu-

tll cnr !;o111llr'llt: -Llail ii\ \\llt!-r(- st'ils' 0t high ''t':'rnic lll'rltt-r

conlillrlin! aLlillPrr\i\ u lltrt- tht phr:tcll 'rtttl ntllrliiilllui

conditiirn: i,r r1ticr,,bitrl -{i()\\lh (11n1()I be col1iPlLrteiv contrtilled.

aniJ a largclr Ltnknolrn tllt.ittbt.tl rrrll'lnlunit) i\ present. Eranl-

製ilcul」ねFli:だ11lliF[Ft札獣W‖llnl猟轟Ia鍵

附f凛 1薯 11風)lin」責1撃。∴篤H∬蹴∬t選尊plicationsヽ 11l bc discusscd in this sCCtiOn

31ode9radation Using Natural MiCrobialCorllmunities

IPIittF‖1棚新ざTTi朧瀞ぶ緊i諄寝黒:

prよ iou、 l、 (｀('('/'59イ ).nlincralization i、 lhc llanSfOHllation o「

()rgを llliC nlolcculc、 lo 11111lC'cti l()1■ ■ヽC()litillall、

itヽ tlS assu11lcd gi、 cn tinlc and thC a11110st inl｀initc

ェI:や|1鍵:1罵gギ寵:キ1llti11:|=ポ ilまま:1属li鷺:lT

Oい、cい ltiい n、 。f na■ 出 i mdヽ )nthctic orgtti… ■甲 ■nd accu―

1lRliatioll i1l ntlttll.llビ l)ヽ'il()nnlclltヽ .hOヽ cヽヽ cr 卜c=an lo ralヽ CЧヒIC｀ ~

ti()n、 ュ1,o tlt[l〕 c tibilit卜 ol｀ nlicroo「=ani｀

111｀ 10 dCgratlc(hescヽ tll ic(1

、ub、 ltHlccS tlll(l thC lolc()i thC c11ヽ il ollnlclnt{CItl卜 、 allaCrobic c● :l「

‖1墓1驚[‖ili鮮湛‖i鮒黒1灘‖淋l醐i鸞肺=思

(‖

織:I里‖謝ド(濯#寵l:起II‖ ll。 lliふ「llよ1よ

週lIIli椰‖IIt卜iぎ1織IIIII11螢 1戦[liヽ c(11「o lllヽ aヽtcrl othCl tヽtldic、 llndicatc th.lt h)drofcn cti11卜 c thc

OヽtllCe Of lctluctを ll]t i10r thc dChalolcllation of(liffclcl■ C111011-

1111ダ llII常 i′)|‖;1淵 11ッ‡||′ }‖lぶ オ ヽ`器

下|)力l

Hunnic iCid、 .br。、Inish polynncric lCSiducs t)f lignill dcconl

驀 輔 幕 鮒

md:鰹 °

鷲

JttlHc醍崚 mmm囃 ぃ rC mmJd測

dc gi atlati(), ol thc lllを li 11 、(1■ lCtlll C 01'nl(ln)pc、 tiCidC、 tlin、 l ollnC:

i鼻彙!1熙榊‖illttIIiI瀧iII珪

Chapter 42 lndustrial Microbiology and Biotechnology 4218

' 42.13 Biodegradation Has Several

ngs. Biodegradatit)lt is a iernl that can

I to cle scribe thre e rna.jor t1 pe s of changcs

rlecule. (a) A minor change in the func-

lroLrps altached to an organtc cot-t'tporrnd.

substitution of a h1'tiroxyI gloup filr a

e rroup. (b).Art :r, iulll brr:aking trf ihc or'

onriround inio orgattic fiagtllellts in sircil

hrrr rhe original rnoiecule could bc reciln-

.i. {c') l-he conrpletc clcgradation of all or

ompor.rnti to rninet lls-

(a) Minor change (dehalogenat;on)

Che ln ca structt,re

O~~l~||~C00H

24-D

G~~CH ~C00H|

α6……… …H6…(b) Fragmentaiicn

G6……… α6…嘲

(c) Mineralization

.ciGO…剛―…翻

Approxinrate time todegrade in soil

3 months

2-3 years

81ocked″ ,era pos tiOn

(b)

42 14 The l′ εrα Ellc・ ct and Biodegrtldation. 1ヽ11lo「

ll di■ cl cllCCS CtlI〕 hを 1、で illl10r CllCCl、 ぃ11 lhC bio(lcgl」 (1-

)1'chenlical、 Thc ″′てィ`′

Cticct iヽ ti:〕 inlPo「 ttl1lt Cヽ そl11lplc

」il)i dcgradablc 2、 1-dic‐ hlor()phcno、 )'acctic acid (2、 4-D,

cxposcd nlcta position oll thc ring dcgrtldc、 111 、cヽ 'Cr[J

(b)rCCalcitrallt 2.4.5-trichlorophcll()入 卜'aCCtic acid(24.5-

thc bloc‐ ked nlcta group can pcr、 1、 [forゝ c aI、

。PPO｀ Cd t。 11lc● 1lh・「

い、iti● n ti〕 ビc(ヽ :l]Pい tilld｀ 1`1l l‐ ごcic―

l ti nluch、 1()、、c「 rtllc ]hc″′`ra effect i、

、hlヽ 、、〔l i1l Agure

hi、 、tcl cぃ chClllical(11「 lcrcllcc i、 lhc rca、 on thatthc cぃ 11]―

nrl)rt lir\\ rt it,-'ilri.i.lr i.1 tll.llJrrl',r1lh.'ntltr ile i1l. iif ill :l.j-l)t.uitlt lt cltlrrr,.r. ir: thi,r11111r Ir-rrttirrn. trll hc llrrg-elr Llcrllrie.l inlL singlr \unrrar lil Llrrlrli:- 1. i.5 1t'ichltlrrlDhctlo\\ ileetic ai'itl.\\rthilri)n\tiill(rrllrrliiriIllr-tlllli)\i1i! l.rri11lltlrjstirltlt.'.lilslirr'\r,\ L-lirl \ cill \. llnrl litu> t: i-r:cd irtt ltrt,:-t.:ltt lrt rtsit e itttttol.

.\l inrpirr.illtlrl:Ir.l ititttlrttltgrttL lrlrl,-lcSterilillr.lrl ts tltc t-ecr.lg-

nitiort thlit IIltn', 01 ihi'etltlllloutlds that atc arltletl to t'llrirott-nreirl\ llrd chiral. (rl Ir()\\ess lls\ nttllL-1r\ lind Illttltledtless.

\ii.r',roILlittirltt\ ()ititt ilttl dcg|ltr-lc (ltll\ (llle i:otttct "i e ttti.l-\tltncC. tire iltll-r i\()ntcr' '," til tctlluirl ltl tlle ttlr itttttllletli. ,{i lca.t

l--it, r,l ht-tl,iii,1t: ltre.hillii. ThLr. rt i: ilillcal lrr ldtl li-re nel'bi-

e itlc irortrcl lhiit l\ c11..ti'. c ltttd lLi.i, de grltdliirl.'. [(uct':li t'.ttdtr's

irer c -ho',i n thut nricrohtal ctrltttitr-tt.titie s irl dillert'nt e nr iLon-

nre nt\ \\ ill cirtgirrcle rjii'te lcnt ellrttltiottlcrs. Chltll-re . itr .ttt irotr-

nrrnt:ll conciililnr airtl nutricltt:iipltiics c:itt rtltet tllc petterns o1

i irilltl l olnt rl.-.! tildlili(rlt\1i.r,;bi.rl .i)nlrllirtilics cl.tltltg.- ihr'ir chltlt teri\tl!\ lll IC

\ll()n\t ro tllt- lrrirlitirrrl,tf iilrlrgellie ',r trrglttlic -iLtllsll-lttcs. [1'l par-

liculel cLrnrptrirrrl. \uch li' l herhicttlc. is.rkler.l rr'Peaicdl\ io a

nriclohirrl crrrrnruttiil. lhe citt-tttttttrtit\ 3dili)ti and fastcr rates l)lilegre.letion i.rrn ()ccur a pr(lcLr\s oi licclinleiiorl (figure 42.15t.

The aiiaptir L' Irr().e\:i o11tn is so ci'1'ectir c tilrrt this cnrichrtlent

culturc-bes.'ti ripprouch. estahlished on tite Principlcs elucidlted

b1 Bcrje rinck \.st,t' p. l/ ) can be Lised to isolate organisnts r"ith a

dcsir.-d se't ol.crpabililies. For erantple. a microbial comntunity

can heconrc so cfficient at rapid herbicide deqraclation that herbi-

cidc cl1-ectii e rtess is dintinislted. To coullteract this 1-rri-'cess. her-

bicitlc. clin he changed i(r tirro\\ thc nricrobial ctttittttunitl.ollblllrn.:. ihrl. i'r i':ar\ ilt! ihc .'ttcitir t'llr'Si o1' tilt cht-rlliceis.

i)t'gi:il:tlt,,rt iri{iar'\.a\ ihat,rcCttt in rtlll. llso clitt L-.c uscd illl:rtSr' :-lric J:,ii.r,l.riiLrtl ir1'hrrirtlcltti,)ll\ ()! \I'3:ta\ ti(illl iigl'tcui-

tLllal opL-ratl(rris. in a tcrltirquc talleii land farnring. The uasle

Microbial Growth in Complex Natural Environrnents 9創4219

，Ｃ●ｏＬｏＯ）

ゆＣ一Ｃ一ＣＥ

Ｏ」０つ一０一０」０エ

Time

Figure 42.'l 5 Repeated Exposure and f)egradation Rate'

.Ariilition ot att herbii:ide to rl soil cln resull in changes ill the

dcgradatir e abiiitl' o1' the nlicrobial cotlltl.ittnitr'' Relative tlegra

ri:rti0n ra{cs lor an herbicide a1'1er initial lddition to a silil. antl af

l('t I('lrc:llc,I c\p{)\tllt lrr IItt rittttc .h'ctii'"rl'

Figure 42.16 lヽicrobial‐ l■ lediated 卜ICtJ Corroslon. Thc

lllicr`ヽ biol()tical con osi()n ofir()niヽ an]lo「 p10blCin Thc=raphiti―

zati()11()fi「 OI〕 undcl a nl、 l blcb()n tl]c IDipC Surll、 cc tl1lo、 1ヽ1liCr001-

gani、 ■、 i1lcluding Dど 、′``rr l〃 '′

ル,toこ ()ロス)dc tlnc inncr sulfacc

The nrethanogens. an inlpo(ant group of the archaea that can produce

metliane . har.e had ri'idcspread impact on the Earth. Dr'\pitc intensir e

research. neq' discoveries are still heing made concerning lhese nli-

croorganisms. Methanogens have nou' been found to cont'ihute to thc

anaerohic corrosir-rn ol st.rft iron. Previousll' the microbial group usu-

ally consiriered the rriajor ctllplit in the anaerobic corr)sion ploce\\

r,,as the renus Dr:-safbi iDrio, lvhich can use sulfate as an oxidant and

h1'cirogen produced in the corroslon process as a reductitnt'

Nlethanrtgerrs cail ilse elemental iron as an electron source in their

metabolism. It appear-s that corro'\ion mev occur even rvithout the

pre -qeircc o1 sulfate. ri hicil is requirerl for functioning of Desullitvib-

no. Rates of iron removal b1 the methanogens are around 79

mg/1.000 cm: ol sutlace area ln a 2':l-ht-'ur period' This rn',u nt't tttn'a.lugh rate. but in ir'lation tl ihe plannetl service life of metal struc-

Iures in muds atrLJ suhl,surl-ace :olls-po:sib11' l'ears and decades-

such corrosion can become a major problem- Continuous ellt)r1s to

improYe protectlon of trtrn structure-t $'ill he require'J in vie $' of the

r-lirer.itr ul tr.'n ., rr, dinr iili( rooT-:.ir:'lll'.

nrate rial is incor.lturated into the soil or allo\.'efl 16 ilirr,r' ltct-riss [he

soi1 sLrrface. u herc rit-gratllttiotl occurs.

It is intportant ttl enrphasizc il)at such degradation Pr(lcLrsses

tlo not alrr a),s reduce en!ironntentnl problerrrs. In I'act. tlle partial

degradation or modif ication of an organic compound rna)' not

lead to decreased toricitl. An exanlple of this process is the rni-

crobial nretabolism ol 1.1, I -trichloro-2.2-bls-(p-chlorophcnl'l)

ethane (DDT). a renobiotic or foreign (chemically s)'nthesiT.ed)

organic cort.tpound. Degradation relnoves a chlorine tunction to

gir e 1. I dichloro-2,2-bls(p-chlorophenyl)ethylene (DDE)' rvhich

is still of environntclttal concern. Another irnporlant exanrple is

the degradation of trichloroethl'lene (TCE). a u'idel1' used solvent'

tf thii is dc-gredcd urtdcr atllerohic conditions. thc dange rotts car

cinogen vinr I chloride can be * nthesized.

■Diodc,Ind(ltioll al、 、、 ご(11l lCti(1 1い 、、itic、 1)rCをld dalllagc、 a11(1 11-

nanciallぃ 、ヽcヽ lヽctttl c■ 11(｀ 1ヽい ll i、 tl l、 11liCtllal l)i1111)。 11を11lt Cヽ anlPIC

Thc l)licI()biallヽ nlcdi(ltctiぜ い1■ o、 it)I〕 ぃ1 1llCtttls i、 I)a]■lcula11卜 ごril―

ictll、 、hcrc iroll pipc、 1lrc tl、 cd il]ヽ tヽitcl l(,g=cd t11lacrobic C11、 iron―

nlc13t、 o: in sccondar)pctF(,lcu131 rccい cヽrヽ ProccSSCS CarriCci c)ut at

oldcr oil llcld、 IIn thcsc r)ldcr llcl(1ヽ 、、をitCr iヽ pu11lpcd doヽ nヽa、 c―

lics of、 こヽll、 to l]orce rc、 idヒ lを11 lDctrい lCtinl t()a CCntral c()llcction

point lf tllc、 'ヽatcr co13taiI]、 lo、 lcヽ cls of organic(1lattcr and sul―

fatc.anacrobic 11licrobial co:1lnnunitics can dc、.clop in l■ lst blcb、 ol

tubcrclcs(figure 42.16,.rc、 ulting in puncturcd iron pipe and loss

of critical ptinlPing PrCSSヒlrc lヽicroぃ rganis111、 that usc clclllcntal

iron as an clcctron donor tiu〔 i11=thc rcdtiction of COc in■ letinano―

gcne、 i、 ha、 c rcccintiヽ いccll di、さいヽ ci ctt (｀ Iicrobial Di、 ersit》

Ecolog}42.2)Bぜ じ(ltl、 せ●fthC、 lヽdき「こ:1=C｀ )「 intcractit)ns that oc―

ctlr bct、 ccn 11licIて 、oritini｀ l11、 tl■ ご l、 iciユ lS thc nccd 〔 dcヽ cl(,Iつ

$tratcgics to dcalヽ iヽth 11licr(ぅ いial c(ヽ 1l oヽ 101l PrOblClll、 1,clticalClr - CHC1 -+ CIIIiC - CH:

一 initial degralation pattern

MO●org甲 1甲 ,W“

'9ut

“CめⅢ籠Sms＼ :船董1蘇"側

Юb

出鍛ぼ＼■||,‐・‐■|'■ |||:

「

h4 = Monitoring wells

:|||

Figure 42.17 A Subsurface I'-ngineerecl Biurenrediution Srstem.iiirtl it: Jro\sil)lr nl()\cnrcnt. \utrie ntr and (l\\ ge r) (ii\ iiir. pero\1dd. ()r

!r'(rLlnci\\ !ter 1(r ])rOnlotr- nrLtre e liiciclrt i-lcgr':rdati0n r.rl'crtntlintinltnts.

涌ヽ /1｀/1｀

/1ヽ

=Contaminants

Itiirrritolin:, lrnd recoverr u,ells rire u\cd t(l nrtrnit,rr the plumeo1h.r ()\\ qL-n rclL'a,qing cornpounds) ar-c added to the soil and

Give alternative deiinitions for the term biodegradation.What is reductiv€ dehalogenation? Descri-re humic acids and

the role they can play in anaerobic degradation processes.

Why is the "mela effect" imporlant for understanding

b iodeg radati on ?

Discuss chirality and its irnportance for understanding degra

dation effects in the environment.What are some of the irrrporlant mrcrobial groups rnvolved in

iron corosion?

Changing Environmental Conditionsto Stimulate Biodegradation

Olten natural nircrohial conrnr,-uiitres *'ill not be abie to calrv outbiodegradaticn proces\e s at a desircrd nitr' due to iilniting pirlsicalor nutritional factors. For e rample. bioctegradation cilien uill be

linrited hr, loq ori,\.!.ll lercl.. Hrdrocarbons. nitrogc-r.r. phospho

rLtq. and othe r ne.'rie tl nLrinari\ :il.r, rnlrr he absent 0r ai arlablco1llr 111 rl11u. flLrr rti.r.. iirLi' rr:rrrtrnt iJia' {rl dr'rr:idllilt)it. In thc.e

alr\c\. rL ls nL'ac\siir v kr tlelclrnine the Iinriting 1ric1()r\. hlred or.t

l-irbiu''r lrnrl S jrelford's la* s. and tircn to sup|lr needrti nr:rtelials

or.nrririilr thc cnr ironnrcnt. i . . .. ,, ',i:, r ,i : . ,

\lr)-\t ()1 tlrc clirlr ctlorts to stinruiate thc dclra(irti,.e iictr\ i-tie: ol' rnie roor-guni:n.ts irrr olveil thc nrodiircation olr \\ aters and

soils br thc uilditron o1'()\\,gen ol nutrienls. calied bioremedia-tion rrnd slroun iri figure -{2.17. Coutact bctuccn tire nricrobcsantl thc subsllate: the pnrpel physical envilonnrcnt- nutr-icnts.

o\\'gen { in nrost clises): and the absencc o1'toxic conrpounds are

critical rn ihis rrranaged process.

OIlen it rs firund that the rddition of easilr melaholized or-genrc rnatter such as qlucose incrcases biodegradatrr-rn ol recalci-lr'ant compounils that are usuallv not used as carbon and ener-et'

sources bv nricroorganisms. This process. temed cometabolism,is frnding rvidespread applications in biodegradation manase-ment. Cornetabolisnr can be carried out by simply adding easilycatabolized orsanic matter such as glucose or cellulose and the

compourrcl to he degradcd to a corlplex nricrobral comrriunitr'.Plants also nral he used to pror,ide the organic nratter. C{lnrdtaholi:rr is irnfor-rlilt in rran\ dilterent birrde-eradation s\ \ie nt\-

‐■理

4

「

4221 Microbial Growth in Complex Natura! Environments 983

弊 C02

Process F unctiorl

Stimulating Hydrocarbon Degradationin Waters and SoilsEr;rericnccs u,ith rtil 'ttjll. rit itlllirla clli ilrttllttt'llls illlt'ttllttthcst principles. \\iltert uotkitlL rrllh tli'llcr:ctl hr,ltLrcrtlhoirt itl

the occan. contllct bet\\ cctt tht ttticrooiLltnisrll. th.' ltr clr,-tcltrl-ton

substrate . antl othcl e sse ntiJl ttLtllicltts illust bt' ttlltitllltitle .l. Ttr

achieve this. |3llg1s t()rlttlitrtll-! lrtllri.nt\ llrld iill 1r1.1)Jrhilic ihrdrocat-bttn stilLrhle') l)icpilrirti()rt lllt',t hecll Lrs.'ti -1-iri. lrthlilri'.1.Itas acceleretcci the tltslitlrtiltt,rl rliiltt-titt ciLti1.. r,tl .iie k. br .:()to'109i . in crttttpat isrrtt ti ith c()ltlt'rrl t,il :ll.k. u ilelr lllt lttltliltottltlnutrient,s \\'L're not lrvai lanic.

A nlljor challcnge ior thr' 1slll1rlirr1r r'..tr 11lr' l:t'"tt \rti,lr''ciil sprll. u,hich occurrctt ilt .'\llr:k:,r irl \llirch l()8(l Scr crltl drt'

tarcnl allploaclles \\er' Li\i'(l t() itt'tt'list bir"lr:!t;r'lrltion Illcrr'inclLtclecl nutt'je nt additions. chtrrticll d j\lt!-!'\ltttt\. hi()\Lll iil.tlltllaclditions. arrri the use of hicit Jrrr'\\tttL- \1..trll T]1c Ll\L'(i1 il rlli-crobiall)' prodrtccd Slr colif id crlrul:i1'i.'i i.trt. irrt'r trr helpiLll

Thesc bictretttetlieltott;t|1ll-i-rltihr'.:il'a ll\!r LlscLl itl 'Lrils lLllti

seditlents. Filreranlpl.'.:i Illlirltlc r\\r'|-\t.l:lc J\l ,rrtr\\ tltn ltt Lt'ed

to degrade PC'Bs in riret .cdittte nt:. Irirst. piiltiltl tlt-llltlogcnilti(rllof the PCIBs occur\ nalurallr urttler ltnrL-r(lt.riL trrnililioils Tilen

the rnuds are aerated 1(l proittolc tl.tt- cotlrplelc de gllttlaiion tlf thc

less chlorinated rcsidtie s ie haptc-r opening figurc r

Stimulating Degradation with PlantsPhytoremediation. or the use ol pl3115 to stinrulate the ertractitln.degradation. adsorptitrtt. stahilization or r.olatilizrttirrn lrl aL)lltllrll

inants is beconlinS an inlponant pill't (r1'hii'tleSr:lJ,rtj,'n rt','htroi-

og1 . .A plant proVidcs nulriL'llt\ th lt '1ll()\\ rrrlllt-i:ih" rlr'|l r\ "t.Lrrin the plant r(rot zonL' or ritizrrrpilere rfigure 11. I E ,. Pllr i,rlcrllcdiatinn lll-\g inClude:1..liint a!tltrilrLtIj\rr.t\ l(),ia"t,1.1.1::r,ri. i:-llitl,rhi

■■

■■||=

こ え

CI CIGOGCI CI

ing t. enhanCed dr:gt-adlrti0n rtf cOntnrrrrntrnt:. Thr tlrgftttllrtion rri'hcrltihlLlt tlbetlzctlt is shilull !: llll Cr:tlllp1t.

Phytoextraclion

Phytodegradation

Rhizofiltration

Phytostabiiization

Phytovolati Iizarion

Llsc 0l pollurlnt accuntul;tting planls to removenrelals or orgrnics iioir soil by concentratitrgthenr in the harvcstebl!r plant pans

I -- | 1'l ril'' :r: 'l r.. .. ii''CJ l' lc'o 'l:alll\m\ Ir'

dcLirrili,rrgliric prll uiants

Lrsc of pllrr i|r)t\ 1(r 3Lr\(lrb uld ad,.orh poilulilnts.

irrrinlr r,rrtnis- 1-r(ri| \i'iiler and equeous rvaste

stIeaJlt \L.\. 01 lllen1s irr reducc lhe bioavarlabilitl'ofpollulants

ln iire e:rr LaorilcrL

Usc ol planr: io r',riatilizc follutar.tts

Based on T. Macek: N'{. Mackova: and J Ktis.

in cnlironmental rcm&lialion. Br.rl.drkrl. .4/lOЮ ExPloitation of plants fc・ r the κmoval of ottanics

18:23-3■ P 25

lizをltion ancI 、:()ltttili/tlti()11 lDl()CCヽ Cヽヽ まヽ :n(ltCd in table 42.12.

Tl a11、 gc:lic PlをlI]ts I1ltl卜 |)C Cl]]ID()卜 Cd i1l Phゝ lorcnnediatioll Usillg

cloning tcchlliЧ tlc、 、ヽlth マ`′ャ,わこれ′ど′ll`′ ll(ヽ どで'ノ'ノ '「

ノゞ, イア9-Sθ、

/10(').thc″7(′ ′へを11ld′ 71て '′ B gcncs htl、ビトccn il〕 tegratcd into a plant

(1′`7/,た

ノ`,ノ

,、 ′` 7/7`ノ f`″ ,て

`)thtIS illをlking it po、 siblc to transforill cx―

tl‐cinCl)ito、 ic()rgallic lllcrcur卜 f()「 11lS tO Clcnncntai nnercucy,v/hich

is icss of an cn、 .ironnlcntal ha′ ard Rcccntly transgcnic tobacco

pla1lts ha、.c bccll cons(1‐uctcd that c、 Prcss tCtlanitrate reductasc,an

CnZ卜 ]1)e fl olll(1ll Cヽ Plい 1ヽ｀ ビーCicfra(Jing btictcrit111l thcrcby cnabling

thc tran、 ビclliti l‐ ]ttll t、 1、 dヽC」「t,tic nilltltc e、 1ご r and nitro aroinatic cx―

PI()｀ 1｀ C｀ ThcゞごInご t:C ti:l卜 i、1ぃ (ililCd IPItull、 rゞO｀ `in s()lutions of cヽ ―

Plい 1`｀

=｀thtttご ぃntI Ⅲl:ヽ itinl｀ Ctilinて tヽ llヽ 1ぜ Itiビ (Dthcr Plants ha、 c bccn

Phytorcmed iation

Chapter 42 lndustrial Microbiology and Biotechnology

Figure 42.'19 Copper I-eaching fronl Lou'-Grade

Ores. Tlte chenri-strr rrnd rnicrobiillosv oi'coppe r ort'

leaching involre ititcrestittl clrllPlelnentlil re:lctjons''l-he nricrobial contributioir is the oridatitln of lenous

iirn ( Fer' ) ttr flrric irln (Fer - ). LtptrtsTtirillrrnt.fe rrrt'tt'i -

,1rltr lutti relaled Inicrrltlrg.rtli:ttlt al't \L'r\ lctire in this

oridatirtn. The terric ion then rcacts cilelllicalll' to sol-

Lri-.ilize thc copper. The solublt coppr:r is reco"ercil t"

l chenrrcal reactiorl uith elcnlenlal inrn' lrhich rcsulis

r n :rn ('lrrIlclitirl .,,|1rq 1' fl rt ;Ii1111,'

r'nginr'elcrl in the s;Ltllc \\l\ to (lt-griille lrichltlr'rt-tht icttc rtl ''lt' i

i-r)n IIlL- rll ;1 I aol-lIll ll I i llllilt i li $ tlrl dlr t tle ctlt.ict'l tl'

Stimulation of fu'letal Bioleaching from MineralsIlittleachin\, ts tlt.. usc of ntic|o()r'gat'risms. u hii.:h pioilLicc rtcids

ironr reducecl sull'ut ctlltlllot-ttl'-ls. lil crcate acidit ctlr itulttlletlt'

that sttlubilizc desir.eil nretal-s lor i-eco\ cr-\" This apprrlaclr is tlst'tl

to reco\,er lnetals front or-cs ltncl nrininq tailings uith in.'tal lcrel:

loo 1ou, 10r ,cinr,lting. Bi0ir:rching carritcl otri bY natulal popLrla-

tiotrs rrf Lt'lttr,.sTtLrillturi-iike specic-s. Thiobtr'rllttr 1/lirrrr'izrlrtrtr'

encl rrlui..tl thiohaciili. l'orerattlple. ullo$,s recoierr of up to 70'i

r)i thc cr)ppci itl lotr gradt' orts. As siltlu n irl figure '12'19' this

urr olr es rhe biololicel ,11id'11ott ot'ctlppe r prL'sellt in lhe se ort-s

to producc solLrble copllcr srrlfate.

It rs apparent tltat italllre $iil assist in blorc-rllediation i1'riVen

a chance. The lolc tlf Illtrul-lli tttict-oorglttlisrlls lll biodcgradation

is nou hetter-apprccirietl. Arl excellent eralrple is thc receni

,,rtrrk.,,,ith the ren ret-slitile 1'ungus Pltattarrtt-Jtat'tt'tltnstt.slttt'

i.rrrir (\[icrotlial l)iversitr' & Ilcologl''{2'3 t

Ol.tenhiodcgr,.lllilrnlndlriodeterirlrationhalt-Ill.ti{)rlle.Jjtire citects. utid it becoiltes intportant til ctlntrol and lirrlit thcse

processr-s b1 e n!irotitlle ntal mallageurent Problenls include ull-

,n ant.,l degraclation of paper. 1et 1'uels. tertiles. and leather goods.

A global colrcern is microbial-based metal corrosiorl'

1. What is cometabolism and why is this important for degrada-

tion processes?

2. What factors must one consider when attempting to stimulate

the microbial degradation of a massive oil spill in a marine en-

vironment?

3. Describe the major types of phytoremediation What is the role

of microorganisms in each of these processes?

4222

Precipitationo{ copper

FeS04

4. How is bioleaching (arried 0ut and \'vhat mi(!-sbiai genera are

invo lved ?

5 What is unique about Fi:aerachaeie chrt'scspltrLtfil? What

does its name mean?

Addition of ttqicroorganisms to COmplexAllicrobial⊂ onlmunities

B(ヽ th i11 1(1い ぃrtit(ヽ 1ヽ alltl liel(1、 t ti(lic、 tittClllI)モ 1`lttヽ こ卜ecil llla(lc lo

、pcc(l t11)c、 i、 tinピ 11licIい bit)lい iゞCtil l)Ioccヽ Cヽヽ 卜〕こ_ldil,ピ kn()｀ 1`1(lC―

ti、 c llnicl otlrtalliヽ 111、 1(ヽ (ヽう ilヽ ｀`[ltclヽ ●l いllncl C()1111〕 :き `、卜ヽ tc:l'｀ [1ヽ

|ボ尋置I1111‖鷲‖》][||:1驀li三:椰1llcnt culttllcs For c、 tinlPIC COnlnlcrcial ctilttilc I)「 CPttratlo11、 arc

tl、 alltlblc to lhcilitatc、 llagc lolnlを 1lion tllnd io i1111,1い C`｀ Cptic tank

PCri'()「 11lallCC

Addition of Mi(roorganisms、″ithout Considering

Protective A/1icrohabitats

Vヽitll thc deヽ cloP1llclnt()1(hC Supcrtう tlゞ

~bゝ=｀ 1`(11lakrabait)in

1 971 tincrcヽ aヽS illitial c、 citc11lcnt dtic lo thc hopc that such tlil illl―

proヽ cd llnicroorganisnl lllight bc ttblc to dcgradc h)'drocarbon

囃 lTR:『 1∫ I:|:書 i::詰∬ 1111tl∫ lattLih∴彗∫‖cC‖

crobc had to sur、「iヽ c and functio1l Enginccrcd 11licroorganisl■ s

、、lcrc addcd toヽ oils andヽ atcrsヽ iヽth thc cヽ Pcctali(♪ n that ratcs Of

dcgradationヽ o■ ld bc、 linlし1latcd aヽ IhC、 cI]]iCIい Orsa1liSnls cstctb―

1lshcd tllcl11、 cI｀ Cヽ (]cncrtlll)such adllitit)nヽ lcd l■ hヽon-lcrnl ln―

crcascs in rate、 ()f thC tlcsircd `ご tiヽ 11) 卜ul i卜1ヽicilil artcr a tc、 、

da、's thc tllicr()bitll ct)11lnltinit)Ic、「t｀ 13｀ Cヽ ｀`きrtヽ ::ミ ,il〔 ir ill trcュ tcd

CuSoo + Feo--> Cuo+ FeSo.

2Fe2｀ S°J3+CuFeS2+2H20+302-'CuSO`+5FeS04+2H2SO`

＼

The basidiomycete Phunenrclutete Ll!.n'sosPori tr? (the scientitlc name

nreans "visible hair. golden spore") is a fungus rvith unusual de-grada

tive capabilities. TIrts organi-rnr is terltred a "i|hile ro1 fullgtrs" because

of it.s ability to degrade li-snin. a randonl-v iinked pltenvlprop'ane baseci

polynrerrc component of u ood (se c .sectittii 28 -l). Thc cellulosic por-

tion ofwood is attacked lo a lesserertent. iesulting iil the charactens

tic white color of the degraded wood. This orsanistn also degrade-s rtruiy amazing range of xenobiotic ctmpound. ,nonbiological foreign

chemicals) using both intraceilular and ertraceliular enzvnles.

As erampies. the iungu-s degradcs hetlzt-nc. toluene. ethviben

zene. and ry'lcnes (the so ciilleil B-l'EX conlporriltls). chlorinated

compounds such as 2.-1.5 trichlorocthl'lene (TCEt. rind trichlorophe-

nols. The latter are present as conl:tlllinants in lr orrd prescl i :ttil e: .intl

also are used as pes{icides. In addjtion. 'ltir':: chlrrl illllied henztilts

can be degraded u'ith oruilhilul 1()iLlene! beitlr plc-r.'nl F't'n tllt ill-secticide H-rrdratnethllnon is degratledl

Ho.'r does thrs ttticrLrtrrganisnt carrr ()tti \uch ttllt:i \1.plre:rtlr

rnosl dcqraaleliott oi'thcse xcnilhiotic alrllln(rurld( (\L.Lir' ,,! .r .ttll'' c

growth. during the seconciarl metaboiic lgnin degradation phase.

l)egradation of some contpounds inr olves important extracellular en-

zymc.s inciutling iignin peroxiti:ise. rnanganese-dependent peroxi-,lase. and glyoxai oxidase. A critical enzlme is pyranose oxidase.

ri hich releases H,O. lor r-rse br, '.hc ntan,uanese-deprndent peroxidase

enz\ rre. The H.O2 also is a piecursor of the highly reactive hydroxyl

radicel. which parlicipates in u'ood de-rradation.,A'ppa,ently the pyra-

nose oxitlase .nzyme is located in the interperiplasmic space of the

tlngal cell rvall. u.'heie it can functrott either as a part of the fungus

or be reicased and penetrate into the ri'ood -rubstrate- It appears thal

th!' nonspecrflc enzll.natic s,vstenl that reiease-' these oxidizing prod-

ucts degrades rnany' cyclic. tromatic. and chlt-rrinated compounds re-

lated to lrgnins.\\'e can e\pect to continue hearing ol manr nerl' acivances in

i.ork gith this organtsnt. Potentjali! valuabie applications being

studicd include grou,th in bioreactors rvheic' intracellular and extra-

icllullr enzlnres can be iraintained in the bioieactor lvhile liquid,.\ i1:r.s fl(\\\ pirst the irrmrtbilized fungi.

litltl cirlllrill \\ siallls. .\1lt'i ltlllllr tlll:il''l't'\'1Lil :1111'lllIL\ li \r' 'i\|,Lrnd thur thc ieck o1 cllectiretla:s ol 'L'lll ltldad ctllltllas riar

rlttr- 1O et least thtee lliat(rr\: ( I I tila llltlrtctirelre:' o1 l.rLrItltlirtr

!l0u n ntictrli)tlillllslll\ a: a foiltl \(rtlrae 1i)l |ladlltol': \Llill it: \\)il

Irrotoz(lir. ilt thc irlehilit\ ()1'tht-\t ltticic-d tllictilrlr!.inl\llt\ llr trill-tiicl tht .onrpouttLi\ to he dt-glitil.'d. lrlil il I the iailLrr.- rrl thc

ltrlilcd n-iicr(tol!attisrtts Ii) \Llr\ ]\ L- :illLl c(rlllllctt' q ith intli-gt-rlilu:

ntieroorglini:nrs. Such a rni-rrlii'icLi tllictoirtg;illlstll tlllir tr.'lts' f illo a()nrllr:1a lutd srirr i"t hccartsc trt'tht lttiilitionltl ilt.'ltt-tti hltr

,1rrr rcqLrirttl t() llllutttltilt thc crillr D\.'\.\tterlllltr hai e bcctl tllaclc ltr tllltkt .it.h l.lhr)l illt)1 \ !rr)\\'ll

.ultuie: rll()ra ailf:Lh]c o1:ulrir.ll ill ll lllttlllill cll\l1r)nlll!-llt h\

Snrrrjpg thcnr itt l(r\\ llutriellt tllttlili t.r sIltliiltL tht ]llitJilrrtullnisnr: be lorr' arldint thcltt ttr iirt L'tt\ il()lltlleilt 'l'ila\e tougllr'ilitlt"Irppr-rilcitts hltr r- itttl-.rpr tci Itlicrobiltl sltrr tr ltl ltlti lllllcli6tt slnle-that. bLtt hr1\c rlol s,,1.'.'ii lilc ilr(lhlL'lll. Ill Ir-ct-lli \clil\. tildrd hli\

i-rt'lr les: intcre\t rrt .itllplr lltldill-! llllrrool!lltti.ltts I(' r'll\ iltrll-nrcitts ,,\ ithout cort:rcitring tht rpt.'illc tlichr ol 1111q11rq'rlr it"llnrL'ntinuhichtlt.-r ltl etostttritc:tiltilirllctiotl. l-llislllislcdtothtlitltj ol natural attenuation. lr hich elllphltsizcs tllt- tl\L- l)l lllttLl-

r ;rl nriclt,bial conrtttutritie. in the tttvtrolltttctltll ll l'lll-I!clllcllt'i r1

poJ I utants.

Addition of Microorganisms ConsideringProtective M icrohabitats\'licroor-canism additions to ttatttral etlr,irotlrrents ciit) be nlore

:uccessful if the nticrorlt sallisnl is arlded together rl itI-t lt illicrtr

hribirat th.t lire> the i)rgani\lll phrsicll P1{rt'cti()ll':is *ell e:

irr:::ihlr \Lrlpl) il'jg ntllllL'lli\. Tili. ill-ik.'. ;i 1lr'::li-'ie ir'r lilr- illi-

-:,,,,1-tLrlr:nr i() \r1r'\i\e in s;ltlc o1 ill.'itltcn.t'a()lllpctitl\a Pic\-

'ul.\ th:,1. a\i\t in lirc lliiiural e ill irtlnttlelit. itlcludrng PrL-ssure

li\,rr llrotoroiln ltrr'(lator\ sllcit lls ciliaie.. lilgellates. alld ail)oc-

bec \lie rolrlLbitlLls ntar ht' eithci livillg rlr inert. i'r'''lrr' ''

I-ir ing Ilicrohabitats. Spccialize-t1 lir iirg nricrol'tlbitats inclr-rde

tlrc sLrrlace'o1 lt s.-crl. I ro(r1. or lt ltal. r',hicll. uith thcir highel nu-

ir icnr 1lu.r rete und thc chlittct iiri in jtial colollizlttion b1'the added

nricr,rorgltnistlts..:ltt I)r{)tcct tllc lldclc'd tllicl'otrt'tl'oln tht'fierc.:a()nrldtiti\ c condiliotls itt thc lllliural ctlr itt,lltlle tit. Eranlplcs ill-ilrrde tht- rtse rrf R/rt:r,lrittrrt liittl lJtir'illrt' iltrttirtgicn.si.t. In ol-der trr

rnsurc thii{ Rhi-t,!tirrnt is itl clo-qe asstlciatirln \\'itll the legtttllc.

scc-rli ltrc co:Lttcl u itlt tltc rtticrtlht Llsills an rlii-t'rrgltrlislll llli\ttll'c.r.tr lllti:rtltiturt is pilrcecl irt tr brttd ulltler tlte 'ccd $'here the ne u iv

tleveloping prinrarr rool rl ill ilalletrlltL'. Itt conlrast' llttcillLt.s

rltut irtt,itrt.sis (flTt i: piaced on the sull'ece ()f the plallt leal'. tlr thc

p1:int is.ncinecrc(i to cottteirt tile tll- gcncs that allou'the plodtrc-

tion of the loric pt'otein in sitl-1. once rt is in-sested. Alier ingestion

br tlic targcl orsanisrtt. tite toxic protern u'il1 be ri'ithin the diges-

ti\. tlrct rrhele it is tttost cllectile. ir ':" '' ', \':, tr i " :ii,,.i.!'iiLIl\ ii:ril I ;l

Inert Nlicrohabitats. Rccentl,t' it ha,s been tbund that nlicroor-ganisrns can be added to natural communities together with pro-

tective inert n'rrcrohabitatslAs an exanlple. if microbes are added

to a soil uilh rnicroporous glass. the sun'il'al o1'added microor-

Sanisnrs can be ntarkedlv erthanced. Other microbes har"e been

t,t.:err etl t() e rartc their ou n ttlicrohabitatsl N'licrrrtlrganislls in

42.24

What factors m;ght iimit the

addition to a soil or water, toability of microcrganisms, alter

be able to Persisr and carry out

Chapter 42 lndustrial Microbiology and Biotechnology

*,ater column orerlying PCB-contarninaied sand-clay' soils

rbeenobsen'edtocreatetheirou'n"clavhtrtches"bl'bindin-es to their outer surface s rvith extrpolysaccharides'

'l'l-resc il-

ations shorv that *ith the application o1'principles ol t-nicro-

eccll0gt, it nla1., be possiblc tO iI()re succcssl.u]il, I.llllnAge

robial cotlli-t'lunitie s in neture'

processes such as changes in bioiilm concentlation gradients. It

is possihle to it-]easure the concentration ol sttbstances from

nranv diffcrent environntents (table 42.13) Aoplications in-

clucle rhe detectiOn ol' ciuci)se. acetic acld. glutarnic acid.

ethaltol. and biochcntical orrsen d'-:tnand In addition. the ap-

plicarion ol biOsensttr.s t() rrer\Lll'e cephaloslr,rrtn. nicoi:rliu

lctil. lind scr,e i'lil B r iilr1liirs hlis bet'n de scrrbed. Ilccentl'v

biosel.;ors har e he en i131'elirpc.1 using inlilluilochenl:ca1-based

detectii)n s\st.ir\. l-htst ner'r biirsensors riill deteci pathog'ns'

herbicides. iorills. pr()tcillS. alrd DNA. \,1anr' of the se biosen.

sors lr. blised on the use o1'a streptavidin birrtln rL-eotnitir)n

svste1ll tTechniques & Applications 42''l)'

i?.epid edr tllaL-s lLre heing nta.lc ln al1 iLre as tlf biosen-qrlr tech-

nologr. The 'c lne 1Ll.1e llrir.lili lllrfril \ clllL.iil\ irl the siabilit)' rnd

.lrr"iril,n o1'lltcse units. r,, iiich are he illg nlaiie tllttre portable attd

sensitir e . \1i.rL,r:rgltlttsttl: allri rtlctllIroljles such ls glucrlsr: can

be ntcttsrtrc.l. ihLr: rllec-ting clititlal ngg{-s irl tllorlern medtcine'

M icrcarraYs

.\ lurt.' L]i11i ,ii tl'r. llt-\r :tllrl dcr ilitllirlg rrlie i obial biole lhn0lrrs-r

tl. l.' 't I)\ \ '(rirl(ll.(\ lll gt'n('llrril\\ lr'lllr'Iiliilt

!-llc t\|1f :'1|il ill r'()llll)]c\ biilltrllclll \\:i1'lll\ tr"'ir'r'llrrlt 1-! (;)

il,. ,"1..'.i iiLl\ il;rLL,. tlrer hltr d r.)r.cL1r1.r,ri lir thi, lire ir ai.L- tha re \uit

(r1 .r \ r!le \\ itt g.-:l,,tlliu.. i'atlol]lirilliilll I)\-\ tt'clln' r1l-'r ' optiur'

Itrtr,i;1.'\.-'rl,r'.irl.riill:i1']'' ''i:'t:'lr't1rrttii"11''I;i|;"'qt'

ing. fhis nticroarrav tecltnologv ila' hccll srtgucslctl to proVici:

thi cquir elL-nt (r1 lhc cilcrlli\t's pcriodic tlrbie lt ol1ers the potetl

trtl ol lis:ltr ills .ili gt-rlcs ttsttl ltl essellli-'le atl ilrganisnl anci cltn

nt,rttit0l crp|r:ssiiltl tll'tcns il1 tltrrLt:ands tl1'gt-'l.ies based tln lhc

pr.incrple:'l,,,.,,, i,, figtrre {2.1 1' ln tilir tt-chnitlrrc' 1 0() to l00 pl

,,,1t,,11e.. cLrtlt:iillillg tlt'sllt'ti \e(lur'il'L-\- llr' sP()l1ed tlnto glltts

-t]r.jt:rli!ltIld]j]lerttllltrct.iltlslrtitltlt.irlri.l.hes.:ltrtlitsarethetltllirlt-l.L l, it}l .|)\'\: 1]ll]11 JtIl( i,\ll|.t.\\](lll rr, r,il. .i/.i1- Birldirlg

o1' tlrt .l)\.\ lrrl \ llliirll\ -gctle: it tlrta'tircri trsine rlpitl photo-

l,r(lll! l;r,'lrit"l ll,: I(i lllllLll'(''

( irtttlltlciltl lllle r()iii-la\ ptLrdltc''t IIIL- ll()"\ ar ailable that con-

tair (t.iJJ iillcll lf.ltllt':1r)l 'LIUL'llllls gcllc c\Prt's-\itllt in '!rrr't'ira-

i ()ii 7\( r,\ ( (,r.(,\ /\i.r(,. F<tr [-- t'ttlr.1.l9(t6pg1 relldrlg llatlrcs ctn Lle

\alillllcLi itl li tltl.toiift.l\ 1Ul ill'11' l(lllt''cllilll'l 'he etltire 1-- r'o1r

desired iunctions?

What types of microhabitats can be used with microorEanisms

when they are added to a complex natural environmenll-

赫hy are`Iants inoculated With θaθア/′υf_1カ

」月βσ′θβSS?

t5 BIOTECHNOLOGICAL APPLICAT!ONS7:roorlianisttts and parls t't ttlicr"()l'Jlrni\lll\' c:pecialll cn-

lcs. ul-L. rrscil rp a ri ide r ariet\ 91'hjote chtl'rlo"1.xl illrpli.ilti()lr5

l1()nitorthe]erelsol.CfitiCalct,rllptlrtlit].itl1i.tc.:ilrl]()Illll!lll, in rrtirttal,s and hullllills. These tccilllitlLtc' hltr c rr ldc a1l1t'1iclr'

ts iJt el]\'iroillllentlrl scicncc. .inirtlai lltttl ltitlttllt.i ilr',tltll. .ltrJ it-'

ic science.

)5ensor5-apidll, tlevelopiilg llr.ll of hiiltechnolocl' 1i111gti t.ll i tltetrsr'

srnational scit'nlif ic irtterest. is that tl1'biosensor proriitctioll'

this lieltl o1' bioclectrorlics' livillg lllicr()organi\ttls l or the ;l

,\'1lres ol orgatleiics) are ltnked s iiil ciecilirtlcs' atld l-'i1rlo:-l

re actiotts ilre converted into clectlicel cttrrttll' bi tllr-'c

)scrrsors (figure 12-20) Biose nsors a|e ltcitlS ilcr c-lopttl t''

rasr.ire sllecilic coltlptlt'tctlts in bt'ct lI ltlIllil' rt POllLtialllt' 1Lr

icct f.iari.r- cr-.r.tp.u'cls in i'o9ii. a1d lo :tur-ir, r'r'rr irirlttllciltlrl

Molecule discriminatingfunction

誓逮癬（ＳｕｂｓｔａｎＣｅ¨ｍｅａｓｕｒｅｄ

E lec t ric

sig nal Biosensors: Potentialand Environmental A

Biomedical, lndustrial,ications

Clinical diagrrosis and biomedical monitoring

,A.gricultura)- honicultural. r'eterinalv analysis

Derection of poliution. and microbial coniamination ot uater

Fermentation :inalr sis and ctrnlrol

\'frniarrino ,rf inrJrr<rrirl oere< 'rnrl lirrrrirtt

RecePtor

gure 42.20 Biosenstlr Design' Bitlseils'rr' 11q- i'rr'l'tr.: rr'-

r:asine applicatiotts in Inedicine' inductrial 1111;1-irlrllrlrr'gr ' riilJ

Receptacle substances(enzyme, antibiotic.

Physical and

chemical change

〓一

J…Bi5tin Bindi and Biotech

Egt u'hite contains man-v proteins and glycoproteins u'ith unitlue prop-

ertie s. (Jne of the most rntcresting. w'hicn binds tcnaciou-sl)' to biotin. *as

isolated in 1 963. This glycoprotein. called avrdin due to its "ar id" brnd-

ing of biotin. *as suggested to pla-v- an tntportant role: making egg rlhitc

antimicrr)bial hy "ryirg up'' the iriotin neeilcd br'rlanv rr,i!r, u,rg'rttisitl:

Ar,idin. uhich fiinctions best under alkaline coniiitiot.ts. has the hishcst

kno',r'n hinding utrnrtl betueen a proteitt anii a lrgard. Scveral Iearslater. scientists at I'iclck & (-o.. Inc. discrrvet'ed n sirttilar protein pro

Cuced b1 an aclinonrlccit. SlrrTr,orini a.s uvirlirti. nhich binds bjotrn at a

neutral pH arrd u hrch dtv:s noi contain carbth)'dri1tes. Thcsc characler-

i,stic> nrakr: \treptavidin'rn ideal biniiing acenl li)rbi()tin- antl it has'r':en

T,arget : Binder

used in an almost unlimited range of applications. as shown in the Box

figure. The streptavidin protein is joined to a probe. W-hen a sample is in-

cubated with the biotinl,lated binder the binder anaches to any available

target molecules. The presence ard location of target molecules can be

determined bv treatinc the sample u'ith a streptavidin probe because the

streptavidin binds to the bitxin on the biotinylated binder. and the probe

is then visuaiized. This derection s)'stem is bein-e etnployed in a wide va-

rietv ol biotechnoic,rgical r:pplicatiorts. including use as a noffad;oactive

probe in hy'briCization studies anc as a cntical component in Diosensors

tbr a u ide range of environi,,cnl:rl ntonitoring and clinical applications.

Not had tbr a protern iionr a "sintple" filamentous bacteriun-r!

Probes

EnzymesRadiolabelsFluorescent agentsChemiluminescent agentsChromophoresHeavy metalsColloidal goldFerritinHemocvaninPhagesMacromcLecular carriersLiposomesSolid suppofts

ｌ

ι

ｒ

Ｊ

AntigensAntibodies

LectinsGlyccconjugates

EnzymesReobptors

nanspod proteinsHydrophobic siies

MembranesNucleic acids. genes

Phages. viruses, bacteria.s,,bcellular organelles. cells.

tissues. whoie organisms

Strept:r\ idin-lliotin BindingSlstems are Finding \1 idespreadApplications in Biotechnologr.\Iedicint., and En\ inrnntentalStudies. lliich IrolccLtie o1- strr-p

iar itlrn. r pr'()tcin de r ir ed il-ottt atr

rlclin()nt\ cctc. irl: lolrr site s brq hich it crn hirld tcnlciousl\ tr) hi-i)till lnolcrl jir lcrir. 13r.rtlaelrilg Ibinrlcr to thc biotin. and a proht-.

such as a f-lLlr)rescent nrolecule. to

tire streptar idin. the target rnolccLlle

can be detected at lo\\' concentra-

tions. Target brndcrs. probes. and

applications are il()ted.

AntibodresAntigensGlycoconjugaiesLecti nsSubstrates, cofactors, inhibitors, etcHormones. effectors, toxins, etc.Vitamins. amino acids, sugars. etc.Lipids. fatty ac,dsLiposomesDNA/FNA probes

All cf the above

―― 乙亀 Z

‰j靴ゴ°百1‖][黒

AP、〈傘oい

°9

∫∫ξＣＯ「０」ＯＱｏ∽

一一０い

ごヽ■ｏ一ＣＯ夕ゝｏ

ゝ＞ｏＦ

T・ 1ご碗IヽL

ξ

∫ で

NSilllIIll[iog町)hy

Reference

Roboticprinting

Computeranalysis

rrit.ie.'fhcst al)ltiotlclle\. brrtil llou lind ill thc 1'ulttrc. tllltkc i1

sihle 1o iirllou lhc t\llrcssi()n ()f thoLlsand\ o1 genes and stuti\

blil rcgLrlatiort ()l' Itticr()biai giru th xl)d resl)onses tu ettr irtrtr

rtei cl'rang.-s.

What are biosensors and how do they detect substances?

What areas are biosensors being used in to assist in chemical

and biological m0nitoring efforts?

Describe streptavidin-biotin systems and how they work. Why

is this technique impoftant?

What is a gene array? What basic techniques are used in this

new procedure?

)pesticides

:re has been a lLlng-tenn interesi in the use olbacteria. fungi.

viruse-s as bioinsecticides and biopesticides (tatrle 42.14t

:se arc dr'firtcil as biological agerlts. such as bacteria. lungi.

t\c\. or thr-ir contponcnts. \\hich can bc used to kill a suscep

e insect. In this stction. ntajor ttses tlf bacteria. lungi. and

rses lo control populations ol.insects rvill be discussed

Reversetranscription

Bacteria

13を lcicritli tiぎ c111｀ incltlに 1ご [1ヽ :llict卜 lli β(′ (′ //``、 1ヽヽせせlC｀ ‐

1)「i1llti「 ll)

β ′/:ll′'71.ゞ

た''7ヽ お (ヽてく'/, マ/2' 1~1li、 bをlctcriulll l、 olll卜 cヽtlkl卜 lo、 ic

t()i11、 cctヽ そ1、 a、 cgctaliヽ c ccll llut cltiriilビ 、Pて

r`ultlli(l13 itiヽ 1()(luCCヽ

al〕 1lntiticclltllar i)1()tCi11モ ぃ、11l cl)、 tal thC Ptlla｀ P01‐ tll卜 :`〉 d)、 that cain

tlct a、 [il、 liCrOhitll il]、 ccticitic F()l 、PCCilic i11、 cct gr■ tip、

1:hC r(lI(1ヽlついrtiI Cr)、 itil altcl c、

1つく)｀ t'rC tO■ lkを lli:le tt)nditi():1、 in

thc llillti=tlt ilttピ l〕 lCI]1ヽ 1● rclcti、 c thcl,「 (ヽ 1い i` ll へitcr tlli、 「cを lCtヽ ヽヽit11

〔l Pl()tC tiヽ C Cnス トnlc tlnc ticti、 ct(ヽ 、ill i、 Prて (ヽlticcd(flglire 42.22, Si、

of thc ticti、 ct()ヽ in tlllit、 intcゞ I alc i1llぃ thC Pla、 11〕 ょ11lC nl卜ranc(lig―

urc 12 22わ ()(ol(■‐nl a hc、 ti=(ヽ nal―、|]をlPc(11)olc tiュ 1● tiピ h thC nlidgLlt

cel;、 a、 、11(ぃ 、、Il in llgtlrc■ 222て / Thi、 lcnd、 lo the l● 、、く,foヽ nl、 )tic‐

balallcc alld Al｀ Il(11ld inall)lo ccH l)i、 1、

Thc nlost rcccllt adヽlallccs in Our ulldcrstandillg tDfお`7(′

〃ιィ∫

′′7“ ′マ′′gノど,7,′ hゞa、 c in、 ol、icci tllc crcation of pest― rcsistant plant、

Thc llrst stcp in thisヽ olkヽvas to inscl■ thC tOヽ 11l gcnc l■ to f (こザ′

This、 ork sh()ヽ cヽd thal tllc c「卜、ttll IDl(■ cin cotlld bc c、 Prc、 、Cd in

tli30thCr()rgarli、 111[1,ld that thc to、 in、、(1、 cffcctil c Thiヽ ■■110r

、ciclltii｀ ic tidヽ tl I]CC ｀`tlヽ 1()llい ヽヽcd i11 19 「ヽ 卜) lhc p:.｀ tit:cti、 I]()f

tonlato plant、 thatご (ヽ IItiincci t11ビ tい 、11l gcllc

B r/7″ ′

'′

7ド た'′ :「ヽヽ ctin bcピ 1(,ヽ 1ヽl il〕 ic「 1lncr]tcrヽ ｀`卜ごFi tilc ccll、

1卜 、e thc sp()rcs tllld Ciゝ 、(al、 ctrc rclctl、 c(l illt()thc incdiunl Thc

Chapter 42 lndustrial Microbiology and Biotechnology 42.26

Test 師ヽ…∞メ凛tミ 遷 纂

奪 たPCB anrplificationpunJ;cation

3

亀:息 :1‐ 3?〔

:,=R=I=二 i

Tt:e Use ofOlder Tech

Bacteria, Viruses, and Fungi As Bioinsecticides: Anwith New Applications

Bactena

Viru-ses

Fungi

Bacillus thuringiensis and BaciLlus popilLiae are the two rna-jor hactena of interest. Bacrllus llitlrirlgietlsls is used

on a s,ide varietv of vegetable and field crops. fiuits. shade trees. and ornamentals. B. popilliae is used primarily against

Japanese beetle lal,ae. Both bacteria are considered harmless io hunrans. Pseudomonas.fluorescetts. u'hich conta;ns the

toxin-producing gene fiorn B. thuringiensis. is used on nlaize io suPPress black cutrvorms.

Three major Yirus lroups tliat do nol appear to replicaie in uirrin blooded aniinals are used: nucleil polyhedrosis l'irus(NPV). granulosis virr.rs (GV). and cr,toplasmic polvhedrosis virus ICPV). These occluded viruses are more protected

in the en\,ironnient.

Or er 500 iiifferent fungi are associated s,ith insecis. lnfcction an,J diselrse occur prirrarilt' through the inseci crtlicle.

Forrrnra_1or9ene., lr"lgheenused.Bar:rr,i,ria btssiattotuttllletarhi:iuntlni.top!iac areusedfbrcontrol of theColoradOpotatobeetle and rhe froghopper ir -su-sarcane piartations. respecliveiv. \;erriciiliun leL:ttnii antl Entontophrhora spp., have been

associateci rvith contrrtl ttf lphids in greenhouse and t-ield environmenls. Coelonrtr:es,:i chytrid. also rs beirrg u:ed ibr control ofniosquitoes.

Toxin binding to

phospho pids and

i∩ seniOn intO membrane

↓NH2 COOH

crystal

Plasmamembrane

Alkaline outcontents

Y

了 躊 鸞 り

/露 獅

側 腕 耐

◇了了鸞扮Protease

+68 l.Da oct're to.ir

--t- ..A.-/ sH ici

t\-t1-

SH

inside ce‖H20,catons

|

iigure 42.22 'Ihe f Iode of -.\ction of the Bat'illus thurinsietr.si.s Torin. (ar Rcleas. o1 the prot()\in fronr the parasprlral btldv and

,iell e: il lr)ss (ri ATP. resultrnc in ceil irttbalrrnce and lrsrs.949

Aggregation andpore formation

H.O cations

ノ

Inside cell

lnside ce‖

Toxin protein Efflux of ATP

ion channel

Outside cell

Microbial Groun lHaior Organisms and Applications

42.28

990 Chapter 42 1ndtlstrial'ν4icrobiO10gy and BiOtechno10gy

p°

電it胤:曜よ」:∵習滞llsPθttltlc｀ uscd Ю COmbtt tC

思模「蹴ittll薫IW[1聯lW撤猛瀑}‖

躍Ⅷittti:嘲 1∬漱1∬]i小鋤dと、

Viruses

鮒鮒III(llttlulぎ :ヽ(1撃ボ∬lCPVヽ alC tinc lCa、 t hOSt Spccillc tiltlscS,[｀

ffccting(1卜 out 200 dil｀

fcrcllt typcs()f inscctヽ ´ヽ1111■11)01ttllll C0111lnCrCiを lヽ 1ヽ1(li pCヽ tiCidC

i、 11larkctCd tllldCr thC trを ldC llalnlc Elcal fOr co11(rol ol｀t1lc cOtt011

b()11ヽVOrnl〃て'″`,r力

′、こ`て

′

(Dnc ol｀ thC ll10ヽ l Cヽ Cll■ 1=〔 1(1｀ そl:lCCヽ 11〕 、ol、 しヽ thc tlヽ C ●1 いtic―

t1lo、:iru、 cヽ thを,t htlヽ c bCCll gcllビ 〔iCtlllヽ 1110dilic(110 PI('(lt,cc ti lつ

て'lCnt

淋 ‖ ギiltttだ ]罵 illll懺il書li繊 |ギ麟 illl

thc l cCO11lbillを lnt bttctll()、 iltlヽ Prc

t()、 ill it tictヽ 1110rC rapi(11ゝ 'thtlll thC l)tl lじllt｀ lrtlヽ 、a11(1 letli tit111■ 1=Clぃ

inscctヽ 1ヽ lllarkcdly dCC,Cascd (|・‐1・ ■11、 ■iヽ ttl ■■t｀ 11 ・

‐ 1

Fungi

Funtiホ o Can bC uscd Ю COnは'in、

cCt PC｀ ｀「

ungalい 1● in、 cdl―

ci(lc、 をlS ll、 lcd ill tablC 42 11、 arC i′ ilndillg lncrCa、 111=tlヽ Cl]n tl=ll~

劇 Imで nedcヽdOpmo■ o「 boPcゞ にdC、 ム PO,C｀ い1腱Ⅲ

dゝ

Aヽ そlilalぅ lc biOill、 cctiCidCs Vヽhich ttrC tlCriヽ Cd llヽ'111 ltingl l11-

cludc ktlヽ tlganlyCillを 11ヽ (lthC l)ol、 0ヽ 11]ヽ t in titlClitioll 1ヽ)CCiを1l111lCro―

bio10=iCJ nndaboliに ヽヽuCh a、 hikkOm、cin and由 CヽⅢnぃ、卜n、 mで

actlVC agaHlst il〕 SCCt｀

The use of nlicroorgall56l-s itl industrial nlicrobiologl' anti

biotechnolo'r,. as rltscussed irl this chaptcr' does ntlt take place in

air elhical ltnd ecological r acuutlt Decisitlils to illlrke a parlicular

product. ,,,,.:1 tilso thc nlethtxjs used' ciin hale iong-teil"-',11:l-

i.,-t -un.r.p..rerj

elTects' as u'ith the apllerir-ance Llf eniiblotlc-

rcsistattl Pathogells lrountl the rr orlcl'

It{rclol.'io}ogv is a ciltical p'irr' of the area of indu:trial ecologl '

concenred *irh trackrng the tlor^'' of elL'nlt'nts atld col'tlpouncis

thoLrgh tht nirturili an't slcial r"orlds' or thc bii;sphere lirld ihe an-

t.l.rrosphcre. \licrobiologr" e-'peciallv ll\ an rtPplied discipline'

siroLrlrl bc .{)rl:irlere(l " itli'l itt stlpporting tt,g111l i"-trrld , :.-

\1iat ()r)rg"illisttls liave bce ll of itnt-ne ll-se berlel-it to llutl-tlttlttr

rl.iri,Lrgh rhc-ir rolc in riiorl prodLrction anj proccssilt 1l: :):l"'

their ir,,.iLrctt to itllpror e hLt'r'lrn lilil anirllal health' in iisrlc''rlttire'

artrl l'Lrt ril.' Iltainte nlttlct- ltrltl itltprilrelnettt tlf ellr ironntcrltal qtlal-

irr. ()tit.'r lttrarr)(rl!ii1ll\lll\' il(i\"c"L'r' ai'c ittiptltlltrlt pathilgetls llrltl

:..i.;: ,;, .1.,',i,,g.' 't'tt' rtlicr.t'r.krgisls hrr'e i.icipcct colltt'irl .r linlit

th.' .teti" liii> ol thc>c- ilartlrliLl tlltctiltlrganlstls''iht {i5i'ilra'1 r lnrl

rr.r. rrl hitri'iiitltl tlllcrilhrlii 1lr'tducl:' sltell as illllihi()iics' hlirc- cotl

trjl.irtcti t,, r, .lciribltlls ol thc hlrtllatl lil'e :lllirl in tile lasl 'clllltl'-\"

..\ ttttct,,lrtLrlrlgtrl ullt' 'iilt-ks ill lttit tlt tht:'' ltrelLs tll'brtl"t'cil-

ilrll(l!\ \iltrrll'1 'tltl:itlt'l 1ll" i'rn'lel tt-f'lt "'tltt'ttt o1'potsiilic tech-

nicll .ieir:i'.ttls"{n trccllcrlt lllil()(itl'tl(ln t() lllc lelationship

he t*ettt tc.lrn{)l{rg} arltl ptrs:ihle sociciltl trl1plicts it Sit.t:'.]t.-'

Satrtr.tcl Fltlrtlllttl t.st't'ttltitr:ntlit l'r' ()Lir-f irst challcrlge' a\ llllcl()-

[]qrl1rg111r. is to titldcr"""t1' tt-t lllLtch lts is ptrssible' the potentiltl

lnlllxtl; ,r1'tlar" lltoiiLl't '\ illl(l P1()ce\\'\ l'rll thc Ittrr'lticl 'r'tit't\ 't:

rre l1;i: \rn iitiar()bi(ril,gt' .i" e ssclllial plll-t oi Lilrs re sprrllsibiliir is

1o h.- .rble ,,' att""t'u'ii*tltte cl'fecttr clrl n ith ihe r ltriilris 'socictll

stekelrrrirrir:.. ltirrr11r the iararetriric urtl rr,rgcr'-tc* rrotc-lrtiril ittl-

f ,,.,. ,,t rrlicrtrbirrl-baseil (rillti trthet ) tcchlltllogie t'

lMPAσS OF ArliCROBIALBIOTECHNOLOGY

讚 mm“

“

ぉ bd∬ edd“ゞ

Briefly describe hOヽヘノthe θaθ 〃′」S r力υr′βθ′θ17S'S toXin klllS

are being used to attempt to conirol in

name for one of these Products?

・踪『謡紫[11=£撃[¶事部照lroadest possible impacts in your discussion of this

Define industrial ecologv'

What are the biosphere and anthrosphere? Why might you

i古 1薫 tiビ term antlЮsphere was COlned?

a particular

terms of the

problem.1

2

3

4.

insects.

What tYPes '

sects? What

Which fungi

of viruses

is a tradepresentlY

2

3

are being used as broPesticides?

42.1 Choosing Microorganisms for lndustrial

MicrobiologY and BiotechnologY

a. Industnal nlicrobioiogr tt;ts hcen LlrL'!i lril ill:ii1Ll

facture such products as anlihirrtl" irllu:1!r

acids. and organic :rcids and has had Illlrl\ ln'l

pc,nunt p,'trtite eflects cltt anin.ial allrl hurllrin

healrh. \Ii'st u'ork in this area has btt-n currietl

out using micrtrorgantrins i'olaled tlom n:llui'

or nlo,iitied hr the u:e rri e lus'il lnutation l"h

niques Brotechnologr inr'rl''e: ihe use of mo-

lecular techoiques rtl trt'ditr lnd imprtlr:

m lL I oUrg -lni \ll I \

h, Finding neu nlicrtrorsanisms in nature lbr use in

lio,".linolngl is a continuing challenge For

nost en\ ironlnalli\. onl\ a ren small part ol th

rrbser' rhle mtcrrlt'ial irrrnmunitr has teen grou'

rtables J2-1 lnd J2'lr' bur malor adranc'-.s t

trtru ing "uniuiiuretl nlitrot'e: are bting nlade

a. i"leation an,l mulatltrn crrntinue to L'e imgrnar

approaches 1-or identi f1 ing nerl m icroorgantsm

Ttrese o etl -.stahlisheil procedures are now bein

4229

conrplertlenlc.l br nloleculltr tcchniqLres. inclLrd

rng ntetabrtlic ettgincet ittg and conlbinatorirl bi

ologl. \\'ith contbinatori:il biologl ltable'12.31.

ir i-. possible 1o tr.in\iet gL'rlr's frolll r)ne t)rganislll

to rlnother oiganism. lnd to lilrnl ne\\'pr()ducts

Li. Sile-directed tlluta!enesis artd prrotein etlgineer

in! arc uscri to rttodill gctte erprr'ssiirn. Tilese

approrrthcs arc' lcadirlg lir lla\\ .tlld L;l-tcn diller-

enl plo.lLtcts riitlt tleir Propcltics l[igure {2.3t'

c. Nalural gartelic (-n-sin.!-r.lnc. is ol inctertsing in-

r.ras1. Thi! i;tr0lres erplCitilll. rlltcrobial re-

ip()n-\c\ l0 slrcrr in lldaptirc nlutitlion arld torccii

!'\olutioir. uith tltr: hope ol idtntilr ing Itticloor

t.!r j.lll \\ rllr L r ir,'t cttr, .

42.2 Microorganism Growth in ControlledE nvironments

Ir \licro,rtgalirmr elitt i.c lrirl n in eonllrl]ltd enr t

1\)nnltnl\ o1 r llitlus t,, pc: :sin-L lirlllglli.:rs lttltl

rrlhtr eLtilttrc' \)'lL'Jr\ ll L['llncr] ctrtlslilttcnts ltle

u.cd. !r()\\th pillllllL'trls alrn be rh(lscll rnd \r jc-d

II tht crrur.e o1'Ir(r\\jllr lt Illicro(llgrttisnl. Thi'

:ririrrr)irail ir Lrt,:ri l,erlrclrllLtlJ lirl thc i-tlllti.liorl rll'

trl,,. l ',:' .r':l rl I:)r-l i'

42.3 Major Products of lndustrialMicrobiologY

:r. \ r iLlt \.ttitl\ ('i..111Ptlttrlrls rle lll'lrlllc.:iJ itl rn-

tjLt.ttirLl rttierohr(rl\)g\ lllllt itllllltcl llttr Iirr'. itl

nrirlr\ \\il\ \ ltable {2'9). Thcrc irriiutlt ltltlbr

,'iL\. alllill(j ll!l!l\. olI:llli. lci(l\. hi(rlloL\rll'l\

\ri.h J\ iitc crelLrrlcrtlitt:. rllLi hio\lilli!aLlllts

rtigures "I2.8. -12.9. rrld J:.1 1) \1icr()olglllll5lll\

rlro crn bc tlscd 3\ hi(rcJlill\\ts t1l cilll\ oilt sl)c

c j1 ic chcttt tcrl 1 !'i].iilrrls r ti gure tl2. I 2 I

,q ;. "1.;;,11'.i,.' .,'1'11', 11r,1 . ,t itt'

Irlr'1ltnl jlilll,li illrlustii:tl 111j;1o[]lrltlgV atld

biotcchi〕 ol●

=ゝ

' ThCSC incltidcヽ ●idcl)' tl、 cd anti―

ttllnor agcnts(table 42.11)

42 4 Microbial Growth in Complex Natural

Environments

a 4ヽicr()orgalli、 11l gro、,tll iI]colnPIC｀ 1](ltural cn、 i

l● ni〕 ,ciltS StiCh tiヽ oヽil、 ti:l llヽ l atClヽ iヽ 11()t tlヽ Cd t()

ciCtiC n3i(｀ l()bial Pro(ltlct、 卜tli to ctlil).Out crlヽ :

「()ni]〕 cntal l)〕 arlagc]〕 ]cnl prOcc、 、cヽ llnCitltlin=

bi()rcnlcdiation Pl`int inoculatioll and()thcl lc―

1(ltcd t,cti、 itic、 11〕 thcsc ctl、 cS tilC nlicrobc、

thclnl、 clヽ.c、 :liC 11()ti'il)aI P!(,dtict、

b Bi(,dCg「 ad(ltio11:s a clitictll Part t)fl■ lttH al、 ゝs―

tcnl、 nlcdiatcd lt、「=cly t,)nlicl o●

「=arli｀

11ls Thi、

C til]inヽ 01、 c nlino「 cha11=c｀ i11を11■ )ヽICCulc rra=´

Incnttltion or nlincializatit,n trlgurc 42 13)

c Bi()dcgratlation ctin bc inl]ticllccd b).inl a n、 i tiC―

t()iヽ inClし 1(111lg o、 )=cl1 1)iCSCl]CC()l ・lい 、cincc htl―

11lic aci(1、 alld ilnc p:c、 clヽ CC 01' I ctiて 1ll) t、 tiblc

organic illattcI Rctlutti、 c dc1lalぃまcnati()ll Pl()

cccは、 hc、 t tlllticl all(tci olpi(: conditi()il、 allti thc

PrC｀ Cncc● 1● l=aniC lユ 1[ittcr Ctlil facilitatc rll()て li「 i―

ct,11()11ぃ |「CCJc tlュ ぅtC():う ,P(ヽ 1l l(1、 il¬ lt lう rぃ CC｀ ｀

ぃi cぃ 1]lcttibい iヽ l il

d Thcヽ tructurc t,1ぃ「fanitこ )`nlP()und、 inllucncc、

dc gitt(ltiti()1]11ビ Ⅲl、 titticiill コIC ilヽ 1)ccilic l● cti―

11)i〕 、()n a 111い 1ビ culc ti、 in lhc′ 11(■ 11〕 )｀ 11()Il l fl=―

1lre 42 141()i il 、alic(1 、1l titttiI[1: :ヽ 1 1llci、 al C

PiCSClう 1 こIc=I atiatiⅢ l, caiュ 1っ c t,「itご :せ て1

C L)C=]tidtiti(ヽ ぃ |]■ l n tl=C■ c ii ctil' 卜c t arlictiい tlt i11

1,lacc 入ヽl〕 cthci tl]、 卜ch=c nl:「 1:1ビ ぃ|、1ぅ

i11、

、ぃil、 o「 l1lc 、ub、u;iatc tflgure 42.17, Such

itil ic、 ctllC Cll)11ヽ 日 tヽittlヽ 1'1ヽ いh C tilcぃ 、こぃ1「うtil―

tiral l,licr(、 bial co11lnl111litic、

「 IDc,「 adatiol]can lc(ld to lll(lca、 cd t()ヽ icil) i11

1]lal]、 ctlSCS Ii l)●t nlal13=cd cを li Cl'tl‖ ) 入ヽidc―

ヽ'ICtld P。

lltlti● :i ctin いCCtll lroll c()「 I()ヽ 1()n i、 a

Questions for Thought and Review 991

particular conccrn、 vith mcthanogcns and Dど ゞ“′―

力1'jわ ″jο playing important Юlcs in this proccss

g Plants can bc uscd tO Simulatc biodcgradation

proces“ S during phメ oFCmCdiation This Can in―

volvc cxtraction. fIItcHng. stabilization. and

、lolatilization oF pollutants([gure 42.18 and

table 42.12)

h 4ヽicroorganisllns can bc atldcd to cnvironincnts

that contain complex nlicrObial communitics

with grcatcr succcss if livitt or inCn mた roh,b

itats arc uscd ThCsc Can incitldc living plant sur―

1■ccs(sCCdS_ roots_ lCa、 cs)or incll matcrials

suで h as nlicroporous gia、s_R力`_r2み

i″″is an im―

portant cxainplc of a lllicnoo瑠aniSm addcd ttl a

COInPICX Cn、 :ronn、 cnt usin=a liヽ ing microhabi―

tat(thC plant rool)

42.5 3iotechnolo9ical ApplKations

a lヽicroorganisnls arc bcing uSCd in a、 vidc ral]gc

of biotcchnological applicatioQヽ stlth as hiく )、 cn―

sOrS(flgure 42_20) 4ヽicroarra)s arc uscd t()

monitor gcnc cxprc、 sion in conlplc、 systcllls

(tigure 42_21,

b BtlctCria 、iruscs and rungi cdll bc tlscd aS bi(う in―

、ecticide、 alnd b:()Pcsticidcs(tabIC 42.141 Bて ,(〃 ‐

/″、 7カ″′ル2マ′′″、′、 iヽ .tn i!111、 lヽtani bi()Pc、 tiCidC

and thc BT=Cnc h(卜 褥 cn inc(ヽ rp)ratcd i:lto co「 n

42.6 impacts of r゛ icrObia1 8:otechnolo9y

a 11ldt:stilai nlicIく )biolo=}and biく itcchnolo=)i cを in

lla、 c loll= tcr1ll and Po、 、ibl)uncxPCCtCd P()、 1-

ti、 c and ilcgati、 c ctl_・ ct、 く)n thC cn、 iroll1llC1lt_

and o:l allill,tilヽ and intllllan、 i:l]Pactcd b)'thc、 c

tcchnologics lヽ llヽ anccsin biotcchnolog)ヽ hOtlld

bc c()Insidcrcd in a b“ )ad cc(,lo=ical and、 ocictal

cて、intc、 1 hヽiCh i、 thC lk)で、1、 of illdtl、 t●al ccolo3).

:rrlaptir e rnut:ttiit:t 9(r9

jnlil !r\phtre 99()

hinirLrlr)entelion 9X'i

lrr()ailisi\ 51 t,)i9

hioJcr:r:id:liott ()J9

bioinsecticides 988

hropeiticide 9Ex-

biopoll rncr 977

bioprospecting 96'ibiortnrediatio;t 982

biosensor 986

hiosplrcre 990

hi0lranstoilrliilioll 97te hir al 9liO

conririnrLlor ial biologt' 966

crrmetabolisrn 981

cortlinurrtts lettl 9l:.tcr nlcntrtion 9l(.)

torced er olution 969

sene rrrt\ 986

industrial ecolog-r' 990

land farming 9E0

lvophiiization 96!rrrr,/r etlect 9ii0

iltetrhLrlic c(rltttol angin!'!'rln! q6--

rlcliib()lic lllth\\ ii\ etlg jtl.':i i illi \1['E I 96r

inieroart rr lechnologr 9s6

Irrt. .,l .rl Lrr.\",ttttJrr,'t"''inirtural atlenualion 93:nrrtural genetic etrginecrinl 969

non-Nervlr-rnian [-.rotlt 97 I

path\\,avxrchitecture 967

phrtorenrediation 9tl3

prinlxrvnielaholite 97,1

proteincn!ineering 9{.6

protopia\l lusion 96-5

rer:rrlcitrancc' 979

reductiredehalogenation 919

regulat()r\ mulanl 976

scaleup 97 I

secondarl rnelabclite 97-l

\enris\,nthetic penicillin 975

site-directed mutasene\is 966

″ ´Hor,, i i it otr:sible lo creete a niche rrr rnicrohahitat

Iilr J Inirrrx)lr'iLn,rill'\\h3{ Jre mc \pc(ul F}inltof concem in tq ing to maLi .ure the microhe can

linci its best piace tc sun ir e ard function lflqrrr miehl the prr'tscnrrmic" era difler fronr

the "senontic era'- l

l. \\-hat further tcchnologicai approacbes u i11 be

requireti to culture n.:icrcrorganistlts that have

nr)t \ et been groB n I Consider the roles of nu-

trient fluxes. comntunication molecules. and

competition.

\\'het nlikc's the rrcri tr1 n:Lutltl seilL'ila !'n!1

nr'er in{ Lrniqlre ) Isn t this :illlpir ri hrl h:: ftct:

:nl t,l , It i t ,r.rlritr .. ,.1 flr. Illlr. llri.'." -: ::

sisrr: $'cre tlrst aLrle to lLrnction l\\'hat are the advanteges o1-microart:ir: for ihe

\tudv ol sene cxpresrion in compler organislli: l

192 Chapter 42 industrialヽ4iCrob101o9y and Bl° teChno10gy

NIost crlnrmercial antibiotic: are prtrdtrced br

oa,,",r"r"aa,aa. lntl onlr rl 1e\\ arc \\nlh!'sl1-ed

i,' i t"*' ,ru orher bacteria From phrsirrlotical

,,na anrlrnnnr"ntal rieupoilts hori tltight vtltl

altetrpt lt) erp)ain this obscrr ltir'n l.

T^" h.r, nruch abotlt lhe henel-iciel u-\'\ ()1 re -

c,r,lrhi,,ant DN'-\ lc'hnLrirrur \\'hi1 alc \()rl'lL- ol

i'n.- ir,rirt",,, lltl''l dislrir entat'5 illiit \h(rukl Irc

coilsitiercti uhen itittlg lriir'r'r1)rc:lili\Ills li)l

lhese aPPiiclitiort: I

8. \Yhr rrright Bacrllrrs tlturingiansis bioinsecti'

citles hc ll interest in otlier areas oi'biotc-chnill-

ug]:': Cnntiat' the nrolecular :lspe'is ()1' tbeil

ttrtlde of aclioll

9 ;,, t.,,, think inlrrnsic biorcnleriiltion tlr":l]]::

[11 ;f out enrirornlental pollutent dL'srrdiltlon

ploblellls l \\.hr trl rr hr nrrt I

lil i\'lut lre si)ll)e ot the pilssible tdrliitllse: oj

bio.scnsiris li-t tlPposed lo lrlore tlltdltional Ph\ 5

ic al atlii chetllic a i llle asur' il ieil t lltoectlure r"'

What are the mluor t1'pes of rnaterials usetl a

nutlients in fcrntentation media'i

In *hll diffe re nl \\ a\ ) can the lerm f ermentailol

be trsed l*nr,-n"rrn,"t"'s can he controlled in a tlotle r

inriustri a1 lernle r.iter'l

;i:,;';; nt,,,.,.r' end sec.ntiar' netabol ites di'

i.,1,,-,,",,r,t o1' thtir srnil-rcsis rlrld fullcli\)rls l

11.

1i

The search for norel plants/rnicrobes and their

.r.Ura,t catl t'c irt Jlrecl ionllr..{ rritli titt erpt'

I*'r..i nun,rnt lt\ noYCl Pitlh')!:( n\ f)i\!rl\s lh(

relative risks antl benefits-are there strategles

,n", ,r" n,nr" likell trl be "rr in-u'itl"'l'rr-r,r.,,rurrw

rtttlitt'l;tratts is a species tlf hacie-

.j, thot i, highlt resisttnt lo radirtiorl' Can rtttt

,i ,,r*-.t ,, '.]lurtthnuln'l'c11 applicatirrn I Horr

*oulil vrlu tesl ii> utilil\'l,'*t in" rrsks rri re leasing genelicrllr rnodi-

tiaJn,ia.oU". or (rncs that are Il()t Ilatural 1o the

Or*tar,,rt ""t'tl;1111si1 \\/hat orecautlt)ns' if

any. would you take? What u'ould be your con-

cerns l4 ;;;. rvhen a n-iicroorganism is removed lrom a

' nuirrui environment and grorvn in the labora-

,".i.*,r, it usually not be able to eifcctively col-

,.ri.. i,. original environment if it is grou n and

"JJ.a fr.r: Cnnsicler the nrture of srou'th me-

dia usetl in the laboralory in comparison to

'rtr)\\tlr cl)nditl{)11\ ll'1 il soil t}r u:ler uhen rt

i.;r;;',,t In urrd'r\lrrrd thts [trndamenlal pr'rh

lem in rnrcrobial ecolcgY'

I lr( n',.1 (lr'rllllr "'" r'" hg"tt dt'' u"' J t'r t

r;t.1 prt, r, i,' .h:rplcl ' "1 trrr h " 'i. (-'irr \ r u

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transsenic Plallls l

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n"an ut"tl l{rgellt'l \\ilh ihe terln indu\t

".ufug" llo\ docs irliciobitl irit)technol()g)

lete to ihesc coltcet tls l

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