Article wjpps 1410169597 2014

www.wjpps.com Vol 3, Issue 9, 2014.

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Saxena et al. World Journal of Pharmacy and Pharmaceutical Sciences

EMERGING ROLE OF FUNGI IN NANOPARTICLE SYNTHESIS AND

THEIR APPLICATIONS

Juhi Saxenaa,b*

, Madan Mohan Sharmaa, Sarika Gupta

b and Abhijeet Singh

a

aDepartment of Biosciences, Manipal University Jaipur,

bDr. B. Lal Institute of

Biotechnology, Malviya Industrial Area Jaipur, India

*Manipal University Jaipur, Dehmi Kalan, Near GVK Toll Plaza, Jaipur-Ajmer Express

Highway, Jaipur, Rajasthan.

ABSTRACT

Nanotechnology research represents a cutting edge technology due to

its diverse applications. The synthesis of nanoparticles with high

monodispersity, specific composition and size is one of the

challenging issues in nanotechnology. In view of this, biosynthesis of

nanoparticle is of considerable importance due to its less toxicity.

Among different biological systems used for synthesis, fungi are

better biogenic agent due to its diversity and better growth control.

Fungi can synthesize nanoparticles both extra and intracellularly.

Mycosynthesized nanoparticles found its vast application in pathogen

detection and control, wound healing, food preservation textile fabrics

and many more. The present review describes fungi as potent

nanofactories, mechanism for synthesis of nanoparticles, characterization as well as their

applications.

Keywords: Fungi, Nanoparticles, Synthesis, Applications.

INTRODUCTION

Nanotechnology, a multidisciplinary science, covers a diverse area of research and

technology in physics, chemistry and biology.[1]

Since its introduction and definition by

Professor Norio Taniguchi in 1974,[2]

more than 90,000 research papers have been

documented in pubmed (till to date). Researchers are immensely interested in nanoparticles

synthesis by physical or chemical means said as engineered nanoparticles (ENPs). Moreover,

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Article Received on

13 July 2014,

Revised on 09 August

2014,

Accepted on 30 August 2014

*Correspondence for

Author

Juhi Saxena

Manipal University Jaipur,

Dehmi Kalan, Near GVK

Toll Plaza, Jaipur-Ajmer

Express Highway, Jaipur,

Rajasthan


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vast applications and rapid utilization of ENPs would inevitably lead to the release of these

materials into the environment and different ecosystems including aquatic and food chain and

hence adversely affect algae, fungi and plants. ENPs penetrate the leaves and enter inside the

cells. Inside cells, ENPs induce the alterations of membranes and other cell structures and

molecules, as well as other protective mechanisms possibly by generating free radicals. In the

past few years, interest in biogenic synthesis of nanoparticles by plant, fungi and bacteria has

been increased as biosynthesized nanoparticles show good polydispersity, dimensions and

stability.[3]

A large number of micro organisms apart from plants have been found to

synthesize nanoparticles, either intra or extracellularly. Nanoparticles having size range

between 0.1-100nm can be attributed to applications in green energy, medicine and

diagnostics, optics, electronics, water treatment systems and even many more in the recent

years.

In this review, we have discussed biogenic approaches for the synthesis of nanoparticles

involving the use of microorganisms such as bacteria, yeast, fungi and plants. Furthermore,

brief overview on potential applications of nanotechnology and nanoparticles in different

fields have been discussed.

Biological synthesis

The biological synthesis of nanoparticles is preferred over physical and chemical means

because of rapid synthesis, better control over size and shape characteristics, less toxicity,

cost-effectiveness and eco-friendly approach.

Nanoparticle synthesis by bacteria: Among microorganisms, the synthesis of nanoparticles

using bacteria has drawn the most attention. Escherichia coli have been used to synthesize

silver nanoparticles (AgNPs).[4]

The synthesized AgNPs were uniformly distributed with an

average size of 50 nm. Furthermore, Gurunathan et al (2009) have altered the parameters

like temperature, pH and concentration of AgNO3 and concluded that nanoparticle size could

be controlled by altering these parameters.[4]

Juibari et al, (2011) explored the potential of

the extremophilic Ureibacillus thermosphaericus in AgNPs synthesis at elevated

temperatures and high silver ion concentrations. Maximum synthesis of AgNPs was achieved

using 0.01 M AgNO3 at 800C.

[5] Pathogenic Gram positive bacteria Staphylococcus aureus

has been used to synthesize AgNPs.[6]

Interestingly, the synthesized AgNPs showed anti-

bacterial activity against methicillin-resistant S. aureus, methicillin-resistant Staphylococcus

epidermidis and Streptococcus pyogenes, whereas only intermediate antimicrobial activity


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was seen against Salmonella typhi and Klebsiella pneumoniae. Biosynthesis of AgNPs by

Bacillus thuringiensis,[7]

Corynebacterium strain SH09,[8]

Bacillus cereus [9]

have been

reported.

In another study, lactic acid bacteria, Lactobacillus spp., Pediococcus pentosaceus,

Enterococcus faecium, and Lactococcus garvieae, were used to produce AgNPs.[10]

These

bacteria were able to produce AgNPs non-enzymatically through the interaction of silver ions

and organic compounds present on the bacterial cell. Better silver recovery on high pH has

also been shown which depicts rapid synthesis of AgNPs by Lactobacillus spp. Sweeney et

al (2004) have used E. coli to synthesize cadmium sulfide (CdS) nanocrystals

intracellularly.[11]

Nanoparticle synthesis by yeast: Silver tolerant yeast strain MKY3 has been used to

synthesize AgNPs which is producing in nanoparticles in large quantities, with simple

downstream processing.[12]

AgNPs of size range between 2-5 nm was synthesized when

challenged with 1 mM soluble silver in the log phase of MKY3 growth. The biosynthesis of

AgNPs and AuNPs by using an extremophilic yeast strain isolated from acid mine drainage

has been reported.[13]

The synthesized AgNPs of average diameter 20nm and AuNPs of

diameter 20 to 100 nm were well dispersed and capped by proteins secreted by yeast.

Biosynthesis of cadmium nanoparticles by using Candida glabrata and Schizosaccharomyce

pombe was reported by Dameron et al (1989).[14]

Biosynthesis of lead sulfide nanoparticles

by the lead resistant marine yeast, Rhodosporidium diobovatum has been reported by

Seshadri et al (2011).[15]

Synthesis of magneto-sensitive nanoparticles by using

Saccharomyces cerevisiae and Cryptococcus humicola has been reported recently.[16]

Nanoparticle synthesis by plant extracts: Green synthesis of nanoparticles by plants is

gaining importance nowadays because of single step biosynthesis process, absence of

toxicants and occurrence of natural capping agents. Singh et al (2013) have reported the cost

effective and environment friendly approach for green synthesis of AuNPs through the

extract of chickpea leaf that act as a reducing agent as well as capping agent.[17]

Synthesis of

Au, Ag, and bimetallic Au core-Ag shell nanoparticles using neem (Azadirachta indica) leaf

broth has been reported by Shankar et al (2004).[18]

The extracellularly synthesized silver

and gold nanoparticles are polydisperse and exhibiting flat, plate like morphology. Rate of

synthesis of Au, Ag, and bimetallic Au core-Ag shell nanoparticles using neem leaf extract

was much faster than those observed using fungi. In another study, rapid, cheap and


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convenient methods to prepare AuNPs from aqueous extract of Mirabilis jalapa flowers has

been reported.[19]

Curry leaf (Murraya koenigii), a well known and potent antioxidant, has been used to

synthesize AgNPs extracellularly.[20]

The synthesis of AgNPs have been shown to be rapid

and particles are of fairly uniform size 10-25 nm and spherical in shape. Further, it was

observed that the increased broth concentration leads augment the rate of reduction, diminish

the particle size and their agglomeration. Synthesis of AgNPs from Cardiospermum

helicacabum leaf extracts has been reported.[21]

They studied the role of extraction

temperature on formation of nanoparticles and found that improved AgNPs synthesis

occurred at 950C (5min) as compared to 65

0C (5min, 10min, 15 min).

Reductant (plant extract) concentration and precursor solution (silver nitrate) has wide impact

on the morphology and the reaction kinetics of nanoparticle synthesis as reported by Khan et

al (2013).[22]

They synthesized AgNPs using an aqueous extract of Pulicaria glutinosa.

Recently, synthesis and characterization of iron oxide nanoparticles from aqueous extracts of

Hordeum vulgare (monocotyledonous) and Rumex acetosa (dicotyledonous) plants has been

documented.[23]

In addition to that, green synthesis of platinum nanoparticles using Diopyros

kaki leaf extract, [24]

AuNPs using Gnidia glauca flower extract,[25]

AuNPs and AgNPs with

clove extract [26]

and Aloe vera plant extract [27]

have been reported.

Nanoparticle synthesis by fungi: Role of fungi as effective nanofactories is catching

attention from the researchers worldwide. Mycogenic route for nanoparticles synthesis has

been well recognized because this totipotent eukaryotic microorganism has several

remarkable features which have been well documented. Fungi can be used as excellent source

of various extracellular enzymes which influences nanoparticle synthesis. There are various

reasons onto which fungi can be chosen as better nano factories over bacteria and plants.

Excellent secretor of protein - Fungi produce large amounts of extracellular enzymes which

catalyse the heavy metal ions and produce nanoparticles. Due to which fungi can produce

Nanoparticle at faster rate than chemical synthesis. [28]

Easy to isolate and culture - Fungi are easy to isolate and subculture as they have simple

nutritional requirements. Serial dilutions, plating and hyphal extraction are the simple


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methods required to isolate fungi. Fungi are totipotent and therefore hyphae or spores can be

used to grow fungus and can be sub-cultured to obtain pure isolate. [28]

Extracellular synthesis of nanoparticle: Fungi can produce nanoparticle extracellularly

which is suitable for easier downstream processing and handling of biomass. Extracellular

synthesis of AgNPs by Aspergillus sp. has been reported. [29]

Better manipulation and growth control: These are additional advantages of fungi to be

used as nanofactories also enzymes secreted by fungi can be used to synthesize nanoparticles

of defined size and shape. Fungi are able to sustain under high agitation and flow pressure as

compared to bacteria and plants. [30]

Strategies for Nanoparticle synthesis by fungi: Top Down: This includes formation of

nanosize material from massive substrate. It involves cutting, etching, grinding by

mechanical, chemical or electrochemical methods (Fig. 1) depending upon the nature of basal

matter.[31]

This may be due to lots of impurities and structural defects in synthesized

nanoparticles by lithography.[ 32]

Fig. 1: Top down approach

Bottom Up: Opposite to top down, bottom approach involves construction of structures by

self or positional assembly into crystals or tubes followed by particle synthesis with

nanoscale dimension (Fig. 2). It is mediated by congregation of substrate to atoms/molecules

and assembly into nanostructures likes nanorods, nanotubes, nanowires or quantum dots. [33]

The key point in mycosynthesis of nanoparticles is the secretory enzymes having reducing


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power that are responsible for reduction of metal compounds into respective nanoparticle. [34]

Bottom up approach for synthesis offers diverse range nanoparticles with more uniformity

and less defects. [35]

The main reason behind this is reduction of Gibbs free energy that results

in synthesis of nanoparticles which are closed to thermodynamic equilibrium. [32]

Fig. 2: Bottom up approach

Mechanism of mycosynthesis of nanoparticle

Fungi can produce nanoparticles both extracellularly as well as intracellularly however the

exact mechanism is not understood completely. Putative mechanisms during intracellular

synthesis include heavy metal binding to fungal cell wall by proteins or enzymes present on it

via electrostatic interactions. Furthermore, the metal ions are reduced by enzymes present in

cell wall. This leads to aggregation of metal ions and formation of nanoparticles. [36]

Extracellular synthesis assumed interaction of metal ions and release of enzyme mainly

reductase with subsequent formation of nanoparticles in solution. [36]

Extracellular synthesis

of nanoparticles has advantages as it does not require lysis of fungal cell, downstream

processing for recovery and purification of nanoparticles [29]

whereas, in case of intracellular

synthesis recovery and purification of nanoparticles from fungi biomass is tedious task and

hence analytical equipments and long processing techniques are required.

The specific role of enzyme alpha NADPH dependent nitrate reductase in AgNPs synthesis

was demonstrated by Kumar et al (2007) [37]

(Fig. 3). The Ag+ ions were reduced by nitrate

reductase leads to formation of silver nanoparticles having 10-20 mm in diameter and

characterized by XRD, TEM, UV-Vis absorbtion.


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Fig. 3: Mechanism of NP synthesis

Further, Srivastava et al (2013) synthesized AgNPs from Halococcus salifodiane and

confirmed the role of intracellular nitrate reductase in it. The synthesized nanoparticles were

50 nm in size as characterized by TEM and XRD analysis. [38]

In addition to that Trichoderma

virens mediated silver nanoparticles production also involves nitrate reductase mediated

silver ion reduction. [39]

Manivasagan et al, 2013 have reported the possible role of nitrate

reductase in silver nanoparticle synthesis as well. They reduced silver ion in culture

supernatant of Nocardiopsis sp. and found extracellular secretion of nitrate reductase

confirmed by FTIR analysis. [40]

The presence of nitrate reductase in cell free extract of

Neuraspora intermedia for AgNPs synthesis has been reported by Hamedi et al, 2014. [41]

The synthesized AgNPs characterized by XRD, FTIR, UV-Visible spectroscopy showed

significant antibacterial activity. In recent times, AgNPs have been synthesized in the

presence of purified enzyme nitrate reductase. [42]

They isolated enzyme from Fusarium

oxysporum on selective medium and then purified by ion exchange chromatography and

ultrafiltration technique. The nitrate reductase mediated synthesis of silver nanoparticle was

dependent on NADPH using gelatin as a capping agent.

Improved surface properties of nanoparticles: Green synthesis is an eco-friendly approach

for generation of nanoparticles with improved surface enhanced Raman scattering (SERS)

properties as reported by Quester et al, 2013. [43]

Authors have conducted experiments to

confirm that biosynthesized metal nanoparticles showed enhance Raman scattering and has

molecules absorbed on metal surfaces. Min et al 2009 reported that silver nanoparticles have

high fraction of surface atoms which shows more antimicrobial effect compared to bulk


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silver. [44]

With similar findings, Das et al, 2009 also reported that gold nanoparticles

synthesized from Rhizopus oryzae have strong adsorption capacity. [45]

Mycosynthesis of different metallic nanoparticles

There have been several reports on extra and intracellular synthesis of nanoparticles by

different fungi using different metals. The recent research on synthesizing nanoparticles using

various fungi is enlisted in table 1.

Silver (Ag): Afreen et al, 2011 have reported extracellular synthesis of monodispersed

AgNPs by Rhizopus stolonifer which is cost effective as well as eco-friendly and

characterized by UV-Vis, SEM, TEM, FTIR and AFM. Further they have also extended their

studies on determining the antibacterial activity against multidrug resistant Psuedomonas

aeruginosa isolated from burnt patients. [46]

Role of Phoma glomerata in the synthesis of

AgNPs has been investigated. [47]

Characterization by UV-Vis, SEM and FTIR characterized

Nanoparticle confirmed capping over AgNPs. In this study they have also suggested that

capping by biomolecules could serve as better candidate for drug delivery system.

Furthermore, they have also investigated the enhanced antibacterial efficacy of synthesized

silver nanoparticles against resistant E. coli, P. aeruginosa, and S. aureus.

Table 1 - Fungi in the synthesis of nanoparticles

Nanoparticle Fungi Size

(nm) Morphology References

Ag R. stolonifer 5–50 Spherical 46

P. glomerata 60–80 Spherical 47

F. oxysporum 50 Spherical 49

F. solani 16.23 Spherical 51

F. solani 3–8 Spherical 52

P. sajor caju 5–50 Spherical 53

A. alternata 20–60 Spherical 55

F. acuminatum 4–50 Spherical 56

P. fellutanum 5–25 Spherical 57

Penicillium

brevicompactum

58.35 ±

17.88 Spherical 58

A. clavatus 10–25 Spherical,

hexagonal 59

A. flavus 17 ± 5.9 Spherical 61

F. oxysporum 20–70 Multishaped 62

V. volvacea 20–150 Spherical,

hexagonal 63

Pestalotia sp. 10–40 Spherical 64

A. clavatus 10–25 Spherical, 59


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hexagonal

Penicillium sp. 10-40 Spherical 67

B. bassiana 36.88–

60.93 Spherical 73

C. keratinophilium 24–51

Spherical 80

V. lecanii 20–50

Spherical 80

F. oxysporum 20–40 Spherical 80

F. oxysporum 300 ± 57 Spherical 69

F. oxysporum 50 Spherical 70

Cylindrocladium

floridanum 25 Spherical 72

Trichoderma sp. 8-60 Spherical 74

Saccharomyces boulardii 3–10 Spherical 75

X. oryzae 14.86 Triangles and

rods 72

Au F. oxysporum 20–40 Multishaped 81

H. solani 2–70

Rods,

triangles,

pentagons,

stars, and

pyramids

85

F. semitectum 18–80 Multishaped 86

R. oryzae 10 Multishaped 45

Penicillium sp. 30–50 Spherical 88

S. rolfsii 25

Triangle,

decahedral,

and

spherical

89

F. oxysporum 34 91

Lentinula edodes 5 to 50 Spherical 75

Neurospora crassa

10-200,

6-23, 3-

12

Triangles,

Hexagons,

Pentagons,

quasi-spheres

43

Penicillium

aurantiogriseum,

Penicillium citrinum,

Penicillium waksmanii,

153.3,

172,

160.1

Spherical 76

A. fumigatus,

A. flavus 17.76-26

Triangles,

Hexagons,

Spherical

77

CdS F. oxysporum 5–20 Spherical 95

Magnetite

Lactobacillus sp.

S. cerevisiae 2.5–5.5 - 96

C. versicolor 100 Spherical 97


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S. pombe 1-1.5 Hexagonal

lattice 3

F. oxysporum 20–50 Quasi-

spherical 99

CdSe F. oxysporum 9–15 Spherical 100

Se A. alternata 30 ± 5 Spherical 101

SrCO3 F. oxysporum - Needle

shaped 102

Si F. oxysporum 5–15 Quasi-

Spherical 103

Ti

F. oxysporum 2−6 Quasi-

Spherical 104


Spherical 103

BaTiO3

Lactobacillus sp. S.

cerevisiae 8–35 - 105


Spherical 104

Bi2O3 F. oxysporum 5–8 Quasi-

Spherical 107

Pt F. oxysporum 20–60 Triangle 108

F. oxysporum 10–50

Triangle,

hexagons,

square,

rectangles

109

F. oxysporum 5-30 - 78

N. crassa 20-110 Spherical 80

Several species of fungi like F. oxysporum, [48-50]

Fusarium solani, [51,52]

Pleurotus sajorcaju,

[53] Fusarium semitectum,

[54] Alternaria alternata,

[55] Fusarium acuminatum,

[56] Penicillium

fellutanum, [57]

Penicillium brevicompactum, [58]

Aspergillus clavatus, [59]

Aspergillus flavus

[60] have been known to synthesize AgNPs. Jain et al, 2010 confirmed the presence of

extracellular protein of molecular weight 32 kDa during synthesis of silver nanoparticle using

cell filterate of A. flavus. [61]

Furthermore, AgNPs were synthesized by exploiting the fungus

F. oxysporum through biotransformation. [62]

Extracellular synthesis of AgNP using extract of

edible mushroom, Volvaniella volvacea as reducing and protecting agent has also been

documented. [63]

Endophytic fungi, Pestalotia sp. isolated from leaves of Syzgium cumini has

been used to produce spherical and polydispersed AgNP having average size of 12.4 nm. [64]

They have reported this silver nanoparticle as better antimicrobial agent by evaluating its

antibacterial activity against S. aureus and S. typhi. Sanghi et al 2009, synthesized protein

capped AgNP using fungus proteins of Coriolus versicolor. [65]

The amino group of protein

was found to be bound on Ag NP as determined by FTIR. The reaction rate was much faster

during NP synthesis under alkaline conditions. Hexagonal and spherical shaped AgNPs were


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synthesized by using A. clavatus isolated from stem tissues of Azardirachta indica. [59]

Trichoderma reesei an eco-friendly fungus known for its large amount of extracellular

enzyme production has been used in the AgNP synthesis as well. [66]

This fungus can be used

for scale up production of AgNP due to its capacity to produce large amount of extracellular

enzyme.

Endophytic fungi living in symbiotic association with plants are also involved in AgNPs

synthesis. The endophytic fungi, Penicillium sp. isolated from Curcuma longa leaves were

found to be excellent producer of silver nanoparticles as reported recently. [67]

Furthermore,

these silver nanoparticles were also found to be effective antimicrobial against E. coli and S.

aureus. In another study Qian et al 2013, synthesized AgNps from an endophytic fungi

Epicoccum nigrum isolated from cambium of Phellodendron amurense. The synthesized

AgNP was found to be highly stable even at varied pH and temperature. [68]

Gold (Au): Being more toxic to the fungus than silver, F. oxysporum mediated AuNP

showed more aggregation and irregularity in shape and size. [81]

Intracellular synthesis of

AuNPs by using Penicillium sp. has been reported by Zhang 2009. [82]

Variation in the

temperature was found to control the size of biosynthesized gold nanoparticles. [82,83]

Shankar et al 2003 synthesised AuNPs using endophytic fungi. Colletotrichum sp. isolated

from leaves of Pelarogonium graveolus as determined by TEM analysis. [84]

Several reports

on mycosynthesis of AuNPs using Helminthosporium solani, [85]

F. semitectum, [86]

Tricothecium sp., [87]

R. oryzae have been well documented. [45]

Rapid extracellular synthesis

of AuNP in cell filterate and intracellular synthesis in fungal biomass by Penicillim sp. has

been performed by Du et al 2011. [88]

They synthesized gold nanoparticle in one minute via

extracellular and intracellular route. Mechanistically role of NADPH dependent enzyme in

AuNPs production has been investigated by Narayanan, 2010. [89]

The Sclerotium rolfsii

mediated gold nanoparticles were found to be spherical and anisotropic that is of variable

shapes as triangle, hexagonal rod and decahedral in shape. Size shape and state of

aggregation of nanoparticle is determined by various factors including various concentrations

of precursor salts, different cellular fractions of culture. Deepa et al 2014 synthesized AuNPs

from culture filterate of F. oxysporum and found diverse shape and size of AuNPs in the

presence of different cellular fractions. Specificity and sensitivity of assay determines the

pathogen detection in less time with more accuracy. [90]

F. oxysporum mediated synthesized

AuNPs and AgNPs conjugated with Candida sp. DNA allowed its rapid detection in modified


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PCR assay. [91]

Synthesis of AuNPs by edible mushroom Pleurotus florida has been

documented. [92]

The synthesized AuNPs showed anticancer activity against human lung

carcinoma (A-549), Human chronic myelogenous leukemia (K562), Human cervix (HeLa)

and human adenocarcinoma mammary gland (MDA-MB) under invitro conditions. Synthesis

of catalytically active AuNPs by P. florida has been reported. [93]

The glucan content of

mushroom was responsible for stability of synthesized AuNPs.

Cadmium (CdTe/ CdS): Synthesis of CdTe quantum dots by using F. oxysporum has been

reported by Syed et al 2013 [94]

and Ahmad 2002. [95]

The nanoparticles synthesized via

biological route shows enhanced antibacterial activity against Gram positive and Gram

negative bacteria. Prasad 2010 has reported synthesis of CdS Nanoparticle using S. cerevisae

as rapid and low cost green method. [96]

Involvement of white rot fungus C. versicolor has

also been well reported. [97]

Other nanoparticles from fungi: In addition to the above, several other metallic

nanoparticles were synthesized using fungi for instances, synthesis of nanosized magnetite by

Mucor javanicus, [98]

F. oxysporum and Verticellum sp., [99]

CdSe quantum dots by F.

oxysporum, [100]

Selenium nanoparticle by A. alternata, [101]

Strontium carbonate crystals by

F. oxysporum, [102]

Silica nanoparticle by F. oxysporum, [103,104]

Titanium nanoparticle by F.

oxysporum, [103]

S. cereviseae, [105]

A. flavus, [106]

Barium titanate nanoparticle by F.

oxysporum, [104]

Bi2O3 nanoparticle by F. oxysporum [107]

and Platinum nanoparticle by F.

oxysporum. [108,109]

Applications of mycosynthesized nanoparticles

Catalysis: Nanoparticles synthesized from fungi have remarkable biocatalytic properties.

Mishra et al, 2014 reported the catalytic power of biosynthesized gold nanoparticles from

Trichoderma. viridae. [110]

This gold nanoparticle in presence of NaBH4 reduced nitrophenol

to 4 aminophenol and also shows antimicrobial property. Further, metal nanoparticles

obtained from fungi can be used in enzyme immobilization for improved enzymatic activity.

Vector control: Role of fungi as antimosquito is well established. Recently Banu et al 2014

synthesised AgNPs from entomopathogenic fungus Beauveria bassiana and found it effective

against dengu vector Aedes aegypti. [72]

In another study Soni et al 2012 evaluated the

adulticidal effect of AgNPs synthesized from Chrysosporium keratinophilium, Verticillium

lecanii and F. oxysporum against filiariasis vector Culex quinquefasciatus. [79]


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Wound healing – Sundaramoorthi et al (2009) have explored the wound healing property

of AgNPs synthesized from Aspergillus niger in experimental rat model (Excision wound

model and thermal wound model). [111]

Their findings suggested AgNPs as better wound

healing property as demonstrated by measuring percentage of wound contraction and period

of epithelialization in dose and time dependent manner. In another study, Phytophthora

infestans synthesized AgNPs was assessed for its wound healing activity. [112]

They found

0.125% (w/w) AgNPs ointments having better wound healing property as compared to

standard silver sulphadiazine ointment.

Textile fabrics – AgNPs synthesized by Lecanicillium lecanii, when incorporated within

cotton fabrics has shown to inhibit the growth of S. aureus and E. coli. [113]

This cotton fabric

cloth can be used to prevent bacterial infections in hospitals. Duran et al (2007) have

demonstrated the incorporation of AgNPs synthesized from F. oxysporum into cotton fabrics

followed by its antibacterial activity against antibacterial activity against S. aureus. [114]

El-

Rafie et al (2009) have shown the reduction in antibacterial activity of AgNPs loaded cotton

fabric against S. aureus and E. coli when washed in laundry after 20 cycles. [115]

Incorporation of a binder in the finishing formulation has retained its antibacterial activity.

Vegetable and Food preservation – Fayaz et al (2009) demonstrated the AgNPs

synthesized from T. viride when incorporated into sodium alginate thin film showed

antibacterial activity and increases the shelf life of carrot and pear as compared to control in

terms of weight loss and soluble protein content. [116]

Molecular Detection: The new PCR assay has been developed by Bansod et al 2013 for

rapid detection of pathogenic fungi Candida sp. from low concentrated DNA. [91]

They have

synthesized gold and silver nanoparticles by F. oxysporum and conjugate these particles with

master mix and DNA sample of Candida sp. This bioconjugate nano PCR assay have shown

high specificity and sensitivity as compared to conventional method.

Anti-bacterial: The number of multi-resistant bacterial strains has been increasing at an

alarming rate and represents a major threat to modern medicine. Emergence of antibiotic

resistance is the consequence of a complex interaction of factors involved in the evolution

and spread of resistance mechanisms. [117]

The sharp increase in antibiotic resistance is caused

by the extensive and improper use of antibiotics in human and animal medicine and in

agriculture. During the past decade, a great potential in nanomedicine has been realized due


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to effectiveness of various nanoconjugates against pathogenic microbes, but they also show

severe toxicity among humans. [118]

The development of nontoxic methods of synthesizing

nanoparticles would be a major step in nanotechnology to allow their application in

nanomedicine. AgNPs have the ability to penetrate the bacterial cell wall and subsequently

damage the cell membrane that leads to death of the cell. Matsumura et al (2003) have

documented the generation of reactive oxygen species (ROS) when AgNPs interact with

respiratory enzymes after entering to the bacterial cells (Fig. 4a). [119]

The generated ROS is

lethal to the cell as it oxidizes all the macromolecules ultimately leads to the cell death. Being

positively charged, Ag+ ions have great affinity towards negative charge present on

phosphate group of DNA (Fig. 4b). Interaction of Ag+ phosphate of DNA inhibits DNA

replication and causes DNA damage that leads to bacterial cell death. [120]

AgNPs have also

shown to modulate the signaling pathway required for cell growth. [121]

AgNPs

dephosphorylate the substrates on tyrosine residues, which lead to inhibition of signal

cascade and thus stops the cell growth (Fig. 4c).

Silver nanoparticles which are spherical in shape synthesized by R. stolonifer have been

shown antibacterial activity against MDR strains of P. aeruginosa isolated from burnt

patients. [46]

The authors have also performed the synergistic effect of silver nanoparticles

with standard antibiotics and found it more effective. In another study P. glomerata mediated

synthesized AgNPs showed remarkable antibacterial activity against E. coli, P. aeruginosa

and S. aureus [47]

and in combination with antibiotics. Pathogens showing resistance to

antibiotics like ampicillin, vancomycin, streptomycin showed susceptibility with silver

nanoparticles alone and when given in combination with antibiotics. Concentration dependent

studies of silver nanoparticles synthesized from endophytic fungi isolated from Curcuma

longa was found to be bactericidal against multi drug resistance E. coli and S. aureus. [67]

In

another study the potentiality of saprophytic fungi, Nigrospora oryzae for synthesis of silver

nanoparticle and its antimicrobial effect was evaluated. [122]

AgNPs were screened against six

bacterial starins namely E. coli, B. cerus, Proteus vulgaris, P. aeruginosa and Micrococus

luteus by well diffusion method and found it bactericidal activity at concentration of

100ug/ml. Several studies including antibacterial effect of mycosynthesised AgNPs against

pathogenic bacteria like S. aureus, S. typhi, E. coli, [56]

Psuedomonas fluorescens, E. coli, [59]

S. aureus, E. coli, [113]

P. aeruginosa, E. coli, S. aureus, [53]

S. aureus, S. typhi. [64]

E. coli,

Agrobacterium tumifaciens, Magnaporthe oryzae [123]

has been reported.


1600


Fig. 4: Mechanisms of antibacterial activity of AgNPs

Anti-fungal: A study designed to assess the efficacy of gold nanoparticles in water

purification was performed by Das et al, 2009 have concluded that AuNP’s synthesized from

R. oryzae are bactericidal against several Gram negative and Gram positive bacteria as well

as inhibits growth of S. cerevisiae and Candida albicans. Mechanistically, the treatment of

AuNPs with fungal cells leads to rupture of cell wall (Fig. 5) as demonstrated by scanning

electron microscope. Furthermore, authors have also found that AuNPs are effective in

cleaning water free from pesticides. In the study conducted to determine the combination

effect of silver nanoparticles with antifungal agent fluconazole against several pathogenic

fungi. [55]

A. alternata mediated synthesis of AgNPs showed antifungal activity against C. albicans, P.

glomerata and Trichoderma sp. In the same study, fluconazole in combination with AgNPs

have been shown maximum inhibition against C. albicans followed by P. glomerata and

Trichoderma sp. In another study role of AgNP in growth inhibition of M. oryzae [123]

and C.

albicans [59]

have been well documented.

Anti-viral: There are several reports available which pictures the effect of fungal

nanoparticles on viruses. Elechiguerra et al (2005) concluded that AgNPs inhibited the

binding of HIV virus to the host cells (Fig. 6). [124]

The nanoparticles having size range

between 1-10 nm attached to the viral surface glycoprotein and thereby prevented its

attachment which is the essential step during virus invasion.


1601


Fig. 5: Mechanism of antifungal activity of AuNPs

Fig. 6: Mechanism of anti-viral activities of AgNPs

Conclusion and Future Directions

A brief description about the fungi in biosynthesis of metallic nanoparticles has been

illustrated. Furthermore, mechanism of synthesis and its diverse applications has been

discussed. Fungi have an upper edge over other biological systems due to its wide diversity,

easy to culture methods, time and cost-effectiveness as well as eco-friendly approach for

nanoparticle synthesis. Myconanotechnology is relatively new development, the future lies in

the optimization of biochemical reactions for producing nanoparticles with improved


1602


composition, size, shape and monodispersity. Genetic engineering technique can be

employed to improve the particle properties in near future.

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