Biotekel-2

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ATMA JAYA YOGYAKARTA UNIVERSITY

BIOTEKNOLOGI

MIKROORGANISME LAUT – 1

Latar Belakang

ZoBell (1946): mikrobia laut kemampuan/

aktivitas penting bg kelangsungan proses

biologi, fisiko-kimia, dan geologi lautan

Mikrob laut jaman modern= industri besar,

nilai ekonomi jutaan $US



Sifat-sifat

1. Morfologi Sel

umumnya Gram negatif (-), bentuk: batang,

spirillum (vibrio), helicoid (kumparan), gerak aktif

dg flagelum (adaptasi), pmbentuk spora (jarang)

2. Fisiologi

mampu mencerna semua senyawa organik &

anorganik (sebag besar), NaCl adl keharusan (air

laut diencerkan s/d 50% pertumbhn terhambat)



3. Kecenderungan Melekat

menempel pd suatu permukaan

biofilm, biofouling

4. Agihan

dipengaruhi oleh: gerakan air laut, jarak

dr pantai, kedalaman, cahaya mthari,

iklim, biotik



Pengaruh Faktor Fisik & Kimia (Abiotik)

1. Suhu

37 oC 42% terbunuh

45 oC 15% (sisa)

optimum= 18 – 22 oC

2. Tekanan

tidak langsung kelarutan senyawa kimia

s/d 3000 atm tahan



3. Salinitas

ion Na+ & Cl- penting

4. Konsentrasi ion-ion (K, Mg, Ca, Mn, Fe, Ni, Co,

dst) kesetimbangan ion, metabolisme beda dg

mikrob darat

5. Senyawa organik

bhn organik terlarut (BOT), bhn organik

partikel (BOP), bhn organik CO2 + KH + as

amino + seny. organik sederhana organisme

heterotrof



Keragaman Mikrobia Laut

Mikrob Laut= semua organisme bersel tunggal

Autotrofik prokaryot (bakteri, cyanobakteri)

Heterotrofik ekaryot (alga, protista)

Virus

Ukuran: 0,3 – 150 μm

3 kelmpk besar*: Bacteria, Archaea, Eukarya

prokaryot ekaryot



*perbedaan berdasar komposisi genetik

molekul ribosom RNA (rRNA): 16S & 18S

a. Prokaryot

Morfologi sederhana: rods (batang), sferis

(mem-bulat), filamen (benang), ukuran 1-2 μm,

kecuali bakteri Epulopiscium fishlesonii

(Gram +, panjang 200-800 μm, hidup dlm usus

ikan di laut Merah)



Sangat beragam dlm hal taksonomi &

metabolisme:

Fototrof

Kemotrof

Litotrof

Organotrof

Atau kombinasi: kemoautotrof, kemolitotrof



b. Jamur

ekaryot heterotrofik jamur benang,

kapang (yeast)

Penting di daerah pantai (sbg dekomposer)

Yeast parasit di laut terbuka (pd

kopepoda)

ekaryot osmotrofik atau sapotrofik



Jejaring Makanan Mikrobia Laut

Produktivitas primer fitoplankton (alga,

diatom, dinoflagellata) kopepoda ikan

“klasik” [peran bakteri heterotrof sbg sumber

makanan bagi hewan benthos]

Botol gelap: O2 berkurang respirasi oleh

mikrobia (bukan kopepoda) Pomeroy

(1974): perubahan cara pandang!



3 hal pokok dlm tulisan Pomeroy:

1) Produsen primer utama bukan hanya

fitoplankton tp jg nanoplankton atau sel-sel

fototrofik (ukuran < 60 μm)

2) Mikrobia (atau organisme < 366 μm) yg mrp

bag dr plankton sumber respirasi & metab

dlm air laut

3) Bhn organik tdk-hidup (terlarut & partikel)

sumber makanan utama bg mikrob heterotrof



Konsep terkait:

Lingkaran mikrobia (microbial loop) oleh

Azam et al. (1983): mikrob heterotrof

(bakteri, bakterivor, protista) brperan sbg

jalur regenerasi nitrogen organik, fosfor,

unsur-unsur bioaktif dan mewakili

mekanisme pengubahan atom C dan enerji

dr jejaring makanan yg berasal dr

fitoplankton



Mikrobia Laut Patogen

Semula diduga mikrob laut tidak patogen!

Contoh: Vibrio cholerae ternyata sumber:

saluran air limbah

Penyakit: tifus, kolera, diare, gastroenteritis

Sanitasi: kesehatan mis. kolam renang



Manfaat Ekonomi Mikrobia Laut

Mis. bakteri

pencemaran: merugikan (biofouling),

menguntungkan (bioremediasi)

Pembusukan makanan: ikan, kerang, udang, dst

(bbrp bakteri tahan thd suhu pembekuan es,

konsentrasi garam laut), bbrp jenis spt dinoflagellata,

diatom menghasilkan racun dlm kerang HAB

Produk fermentasi (menguntungkan): alga coklat,

ikan, cumi oleh Alteromonas, Aspergillus,

Penicillium, Clostridium, Rhodotorula, dst.



Penerapan Bioteknologi

Metabolit sekunder

Simbiosis dg organisme laut lain organisme

manakah yg menghasilkan metabolit sekunder?

antibiotik, racun (TTX, STX)

1947: Rosenfeld & ZoBell bakteri laut memp

aktivitas antibiotik

1966: Burkholder et al. berhasil memurnikan

antibiotik dr bakteri laut



1966: Lovell struktur kimia antibiotik:

highly brominated pyrrole

Senyawa antibiotik = metabolit sekunder

Sangat berbeda dg seny antibiotik yg

dihasilkan oleh mikrobia darat!



Table 1. Marine bacteria that are proved to produce pharmaco-

medical substances

Species/Strain Source Biomedical Activity

Chaina purpurogena sediment Antibiotic, anticancer

Streptomyces tenjimariensis sediment antibiotic

S. griseus sediment antibiotic

S. sioyaensis sediment anticancer

Maduromycete sediment antibiotic

Streptomyces sp sediment antibiotic

Actinomycete sediment antibiotic

Pseudomonas bromoutilis seagrass antibiotic

Chromobacterium sp seawater antibiotic

Alteromonas rubra sponge anticancer

Thermococcus Hydrothermal vents antifungal

Bacillus Deep-sea sediment anticancer

Source: Austin (1988), Jensen & Fenical (1994)



MARINE MICROB BIOTECHNOLOGY

Marine micro represent vast & rich biomedical

resource

Algae, bacteria, fungi can be found in all parts of

the ocean, cosmopolitant, many exist as symbionts

of marine plants & animals, many unculturable

Pharmacological resource potential

Unidentified deep-sea bacterium produce new

chemical class of compounds inhibit tumor cells

proliferation & replication of HIV/AIDS



Several marine fungi produce antibiotics

& neuroactive substances

Industrial resources: bacteria living in

very high temperature water at deep-sea

hydrothermal vent manipulate &

genetically engineered heat resistant

enzymes commercial applications, eg.

Stratagene (California): enzyme for PCR



Bacteria of hydrothermal vent another

potentially attribute: ability to oxydize H2S and

use the energy to fix CO2 organic compounds

Other bacteria: reproduce extremely rapidly

and produce methane biomass conversion

into methane more efficiently

Uncultured microb: genomic & bioinformatic

methods powerful tools to access the gene

products of these uncultured microb.



Uncultured microb: eg. total DNA from environmental sample can be purified without culturing the microb. Environment DNA sequenced analogously to a genome allows access not only to the protein products of uncultured bacterial species but also to the genomic potential of the environment (“ecological genomic”)

Bioinformatic analysis cloning and expression of selected genes from the uncultured bacteria will likely lead to the discovery of novel bioactive molecules!



Uncultured microb:

HTC (High-Through-put-Culturing)produces to isolate cells in very low nutrient mediamicrobial metabolic processes

Use of microtiter dishes and a newly developed procedure in which cell arrays are made on microscope slides and screened by fluorescence in situ hybridization (FISH)



Source: Walsh, 1995



Biomedical Resource

Problem: antibiotic resistant microb

easy to transmit, high prevalence,

multidrug resistant (=transferable

resistance gene), fast resistance

development, malpractice of antibiotics,

etc BIG problem in developing

countries



eg. pathogenic resistant bacteria to human:

oStaphylococcus aureus (MRSA), vancomycin-resistant

Enterococcus faecium (VREF)

oGentamicin-resistant E. faecalis

oTrovafloxacin & ciprofloxacin-resistant S. aureus

oPseudomonas, Salmonella, Shigella, Campylobacter,

Mycobacterium

eg. less available of anti-micotyc drugs: Candida

albicans, Apergillus fumigatus, Cryptocarrion

neoformans, Pneumocystis carinii

Marine microbes = alternative antibiotics!!!



Tabel 1. Sumber bakteri penghasil antibakteri

Sumber Jumlah strain dari Genus

Pseudomonas/

Alteromonas

Vibrio Flavobacterium

Air laut 12 - 1

sedimen 1 - -

fitoplankton 5 5 -

zooplankton 10 1 -

sponge 2 - -

Total %-ase 81 16,3 2,7

Sumber: Nair & Simidu, 1987


ATMA JAYA YOGYAKARTA UNIVERSITYTabel 2. Jenis Bakteri laut Penghasil antibiotik

Bakteri Antibiotik Pustaka

Bacillus cereus QN03323 Tiopeptide Nagai et al. 2003

Bacillus sp. MK-PNG-276A Cyclic decapetide (Loloatin A, B, C,

D)

Gerard et al. 1999

Bacillus sp. PP19-H3 macrolactins Sano et al. 2001

Bacillus sp strain NM12 Siderophore (MW < 5 kDa) Sugita et al. 1998

Actinomycete isolat CNB-632 Marinone, debromo-marinone,

sesquiterpenoid, naphthoquinone

Pathirana et al. 1992

Actinomycete (Chainia

purpurogena SS-228)

benzanthraquinone Kitahara et al. 1975;

Okazaki et al. 1975

Myxobacterium (Haliangium

luteum)

haliangicin Yamanaka et al. 2001;

Kundim et al. 2003

Streptomyces sp Δ-indomycinone Biabani et al. 1997

Streptomyces sp MT-MA 190 lerneomides Capon et al. 2000

Streptomycete Dihydrophencomycin methyl

esther

Pusecker et al. 1997

S. rishiriensies MJ 773-88K4 lactonamycin Matsumoto et al. 1999

S. tenjimariensis SS-939 Istamycins A & B Okami et al. 1979

S. maritimus Enterosin, wailupemycins Piel et al. 2000



SEKIAN

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TERIMA KASIH

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