BIOSYNTHESIS OF METAL NANOPARTICLES BY MICROORGANISMS

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1 BIOSYNTHESIS OF METAL NANOPARTICLES BY MICROORGANISMS Saeid Talebi a,f. Ramezani b,m. Ramezani c Abstract: a; Islamic Azad University,Sari Branch.E.mail:Sa_talebi@yahoo.com b; Shahid beheshti university,iran.e.mail:fr_750@yahoo.co c; Tarbiat modarres university,iran.e.mail:mo.ramezani@yahoo.com Corresponding author: Fatemeh ramezani Due to unusual optical, chemical and electrical properties of nanoparticles, making of them is very important. Nanoparticles are used in many respects in the biotechnology and industry, for example in the fields of catalysts, optics, life sciences, pharmacy, medicine, mechanics, magnetism and energy science. Variety of techniques for making of metal nanoparticles are chemical recovery using regenerative materials, aerosol technique, electrochemical deposition, photochemical recovery, laser exposure and.... But as the term of green nanotechnology has emerged that a lot of attention has attracted and includes a wide range of processes that reduce or eliminate toxic substances to restore environment. Green Nanotechnology also seek more effective alternatives for energy production (e.g. solar and fuel cells). In green nanotechnology, for the synthesis of nanoparticles microorganisms are used. Well known that many microorganisms, aggregate inorganic material within or outside the cell to form nanoparticles. For example, single cell organisms like magnetic bacteria produce magnetic nanoparticles and Dyatumeh produce nano-particles of quartz and other bacteria, viruses, fungi and yeast produce various kind of nanoparticles i.e silver, gold, CdS, Bismut, TiO 2, SiO 2 and so on. In this article we research about the use of microorganisms in the synthesis of metal nanoparticles. Keywords: green nanotechnology, metal nanoparticles, microorganisms. Introduction: Nanoparticles are usually 1 to 100 nm in each spatial dimension considered as building blocks of the next generation of optoelectronics, electronics, and various chemical and biochemical sensors. Synthesis of nanoparticles with different compositions, sizes, shapes and controlled dispersity is an important aspect of nanotechnology. In order to synthesize noble metal nanoparticles specific methodologies have been formulated [1]. Despite the physical and chemical techniques being able to produce large quantities of nanoparticles with a defined size and shape in a relatively short time they are complicated, outdated, costly, and inefficient and produce hazardous toxic wastes that are harmful [2]. Biosynthesis is considered better than chemical and physical synthesis because: 1. Use of expensive and toxic chemicals is eliminated [2] and It is a clean and eco-friendly method [3,4]. 2. Biological nanoparticle synthesis would have greater commercial viability and large savings in reductant and energy costs and high production rate in comparison with conventional methods [3]. 3. Large-scale production by chemical and physical methods usually results in particles larger than several micrometers while the biological synthesis can be successfully used for production of small nanoparticles in large-scales operations [3]. 4. Physical methods need high temperature and chemical methods need high pressure which is a harder situation to provide [3]. 5. Prokaryotes can be easily modified using genetic engineering techniques for over expression of specific enzymes, apart from the

2 ease of handling [5]. 6. The particles generated biologically have higher catalytic reactivity, greater specific surface area [2]. 7. In almost all chemical nanoparticle synthesis methods, a stabilizer is necessary to prevent the aggregation of fine particles. Conversely, in TEM images of nanoparticle samples synthesized with biological methods, it is clear that even aggregated nanoparticles don t have direct contact with one another. This is due to the fact that nanoparticles are stabilized in solution by capping proteins, which are secreted from microorganisms. One important enzyme that may be responsible for this is Cytochrome C [3]. 1. USE OF BACTERIA 1.1. Pseudomonas: Silver-based single nanocrystals in periplasmic space of the bacterium were produced by Pseudomonas stutzeri AG259, a silver mine bacterium (figure 1) [5,6,7]. P. stutzeri also aerobically possessed the ability to reduce selenite into insoluble elemental selenium [8]. Recently, cell supernatant of P. aeruginosa was used for the reduction of gold ions resulting in extracellular biosynthesis of gold nanoparticles [7,9] Magnetotactic bacteria: Magnetotactic bacteria such as Magn- etospirillum magneticum produce two types of particles; magnetic (Fe 3 O 4 ) and greigite (Fe 3 S 4 ) nanoparticles [7]. Aquaspirillum magnetotacticum, produced crystals of ordered single-domain magnetite (Fe 3 O 4 ) particles. Magnetite nanoparticles formed by bacteria such as A. magnetotacticum, Magnetotactic bacterium MV-1, Sulfatereducing bacteria, M. magnetotacticum and M. gryphiswaldense showed predominant morphologies of octahedral prism, parallelepiped, cubo-octahedral and hexagonal prism in the size range of nm[1] 1.3. sulphate- reducing bacterium: In the presence of exogenous electron donor, sulphate- reducing bacterium Desulfovibrio desulfuricans NCIMB 8307 has been shown to be synthesizing palladium nanoparticles on the surface of cells and also can bioreduce selenite to selenium both inside and outside the cell with various morphologies. Heterotrophic sulfate-reducing bacterial (SRB) enrichment from a gold mine was used to destabilize gold (I)-thiosulfate complex to elemental gold [7]. Also these species can form ZnS particles with a diameter of 2 5 nm [6,10]. In that way, Desulfosporosinus sp., a Gram-positive sulfate-reducing microbe isolated from sediments when incubated with mobile hexavalent uranium U (VI) reduced to tetravalent uranium U (IV) which precipitated uraninite were in the size range of nm [1] Lactobacillus: Lactobacillus sp. can synthesis spherical aggregates of TiO 2 nanoparticles [11]. These titanium nanoparticles were lighter in weight and high resistance to corrosion [1]. Common Lactobacillus strains found in buttermilk assisted the growth of microscopic gold, silver, and gold-silver alloy crystals of well-defined morphology [7,12] within the cell with no disturbance in its viability. Gold crystals were found as nm in hexagonal and triangular shapes and as clusters of 100 nm [1] Fig 1.Crystal topologies by P.stutzeri AG259.(a,b) Triangular, hexagonal and spherical Ag-NPs. found at different cellular binding sites[2].

3 1.5. Bacillus: Bacillus subtilis 168 is able to reduce Au 3+ ions to produce octahedral gold particles of nanoscale dimensions (5 25 nm) [6,13] Recently, an airborne Bacillus sp. was also found to reduce Ag + ions to Ag 0. This bacterium accumulated metallic silver of 5 15 nm in size in the periplasmic space of the cell, the culture supernatant of B.licheniformis was used for the extracellular synthesis of silver nanoparticles of 50 nm [1] Fe (III) reducing bacterium: In Fe (III) reducing bacterium, Geobacter ferrireducens, gold was precipitated intracellularly in periplasmic space [1]. Shewanella algae reduced Au +3 ions to nm gold nanoparticles [14] and can produce platinum nanoparticles (see fig 2) [15]. Magnetic octahedral nanoparticles of sizes below 12 nm are formed extracellularly on the surface of the thermophilic ironreducing bacterial strain Thermoanaerobacter ethanolicus (TOR-39) [16,19] Clostridium: within 24 48h after the addition of CdCl 2 and 0.05% cysteine hydrochloride, Clostridium thermoaceticum in late exponential- to early-stationary phase precipitated bright yellow CdS crystals on the surfaces of the cells as well as in the medium [1, 6,17] Staphylococcus: When the S.aureus was subjected to AgNO 3, the reaction started within a few minutes and the color of the solution turned to yellowish brown, indicating the formation of AgNPs extracellularly [18] Cyanobacterium: Interaction of Plectonema boryanum UTEX 485 with aqueous gold(iii) chloride initially promoted the precipitation of nanoparticles of amorphous gold(i) sulfide at the cell walls, and finally deposited metallic gold in the form of octahedral (111) platelets ( 10 nm to 6 µm) near cell surfaces and in solutions [7]. The addition of PtCl 4 to P.boryanum UTEX 485 culture promoted the precipitation of Pt (II) organic material as amorphous spherical nanoparticles ( 0.3 µm) in solutions and dispersed nanoparticles within bacterial cells [20] Entrobacteriacea: The culture supernatants of Enterobacteriaceae (Klebsiella pneumonia, E. coli and Enterobacter cloacae) rapidly synthesized silver nanoparticles by reducing Ag + to Ag 0 [25]. E. coli DH5α mediated bioreduction of chloroauric acid to Au 0 nanoparticles has been reported recently. The accumulated particles on the cell surface were mostly spherical with little other morphologies of triangles and quasihexagons [1]. Cultivating E. coli and a strain of Klebsiella pneumoniae (formerly K.aerogenes) in the presence of CdCl 2 and Na 2 S results in the intracellular formation of CdS [16,20, 21]. A facultative anaerobic bacterium, Enterobacter cloacea also can bioreduce selenite to selenium both inside and outside aggregates[1] Photosynthetic bacterium: Dried cells of Corynebacterium sp. SH09 produced silver nanoparticles on the cell wall in the size range of nm with diamine silver complex [Ag(NH 3 ) 2 ] + [22]. Stenotrophomonas maltophilia SELTE02, showed promising transformation of selenite (SeO 2 3 ) to elemental selenium (Se 0 ) accumulating selenium granules either in the cell cytoplasm or in the extracellular space [1]. S. maltophilia can synthesis gold nanoparticles [23]. In analogous, Rhodobacter sphaeroides produced extracellularly FCC structured lead sulfide (PbS) nanoparticles of size 10.5± 0.15 nm with monodispersed spherical morphology. ZnS nanoparticles were synthesized by immobilized R.sphaeroides [17]. A simple route for the synthesis of cadmium sulfide nanoparticles by photosynthetic bacteria Rhodopseudomonas palustris has been demonstrated [20,34]. Rhodopseudomonas capsulata successfully produce gold nanoparticles with different sizes and shapes [24].

4 2. YEAST: It has long been recognized that among the eukaryotes, yeasts are explored mostly in the biosynthesis of the semiconductor nanoparticles. Peptide-coated nanoparticles derived from yeast species are analogous to the semiconductor quantum dots studied in solid-state physics. But the yeast synthesis mechanism overcomes one of the biggest disadvantages of physical and chemical synthetic pathways: the agglomeration into larger particles. Nanocrystals derived from yeast are naturally stabilized by the phytochelatin layer, which also effectively controls the particle size [16]. Yeast derived CdS nanoparticles show three important features for the quality of colloidal nanoparticles in biological applications: Crystallinity of the particles, narrow size distribution, and good water solubility. Exposure of Candida glabrata and Schizosaccharomyces pombe to Cd + 2 ions leads to the intracellular formation of CdS quantum dots [6,20,25]. Recent studies have revealed the extracellular formation of silver nanoparticles in the size range of 2 5 nm by the silver-tolerant yeast strain MKY3 [26,16]. Torulopsis sp., is capable of synthesizing PbS nanocrystals intracellularly, when challenged with Pb + 2. Crystallites, which are extracted from the biomass by freeze thawing, are 2 5 nm in size [6]. Recently, yeast strains have been identified for their ability to produce gold nanoparticles, whereby controlling growth and other cellular activities controlled size and shape of the nanoparticles was achieved [7]. 3. BIOSYNTHESIS OF NANOPARTICLES BY FUNGI: Myconanotechnology is a new term that is defined as the fabrication of nanoparticles by fungi and their subsequent application, particularly in medicine. Fungi have a number of advantages for nanoparticle synthesis compared with other organisms, particularly as they are relatively easy to isolate and culture, and secrete large amounts of extracellular enzymes, and they have wide range and diversity. Many of the proteins secreted by fungi are capable of hydrolyzing metal ions quickly and through non-hazardous processes. In addition, nanoparticles of high monodispersity and dimensions can be obtained from fungi [27,36]. Shift from bacteria to fungi as a means of developing natural nano-factories has the added advantage that downstream processing and handling of the biomass would be much simpler. Compared to bacterial fermentations, fungal broths can be easily filtered by filter press of similar simple equipment [25]. Verticillium, when exposed to aqueous AgNO 3, caused the reduction of the metal ions and formation of silver nanoparticles of about 25 nm diameters [5]. Verticillium luteoalbum produced gold nanoparticles in spherical and rod shapes [1]. F. oxysporum extracellularly synthesized various nanoparticles like gold [7], silver [5,28,37], bimetallic Au Ag alloy, silica, titania [6], zirconia [7], magnetite, Bi 2 O 3 [7], platinum nanoparticles [35,33,37], Cadmium Carbonate [38,40], Cadmium sulfide [27,38], Strontium Carbonate [39] Crystals in aqueous media. Silver nanoparticles have also been reported from Trichoderma asperellum and Penicillium sp. Recently, was showed that the use of Aspergillus niger [27], Aspergillus flavus [7,12] and Aspergillus fumigates [29] resulted in the accumulation of silver nanoparticles on the surface of their cell wall. Filamentous fungus Hormoconis resinae also can synthesis silver nanoparticles [30]. 4. USE OF ACTINOMYCETES: Actinomycetes are microorganisms that share important characteristics of fungi and prokaryotes such as bacteria [25]. Thermomonospora sp. when exposed to gold ions reduced the metal ions extracellularly, yielding gold nanoparticles with a much improved polydispersity [7,31]. The alkalotolerant actinomycetes

5 Rhodococcus sp. produce intracellular golden nanoparticles concentrated on the plasma membrane [16]. Ahmad et al. reported an alkalotolerent actinnomycetes (Rhodococcus sp) capable of synthesizing gold nanoparticles of the dimension 5-15 nm with good monodispersity formed on the cell wall as well as on the cytoplasmic membrane [4]. 5. VIRUS MEDIATED BIOSYNTHESIS OF NANOPARTICLES Interestingly, tobacco mosaic virus (TMV) was used as template for the synthesis of iron oxides by oxidative hydrolysis, co-crystallization of CdS and PbS, and the synthesis of SiO 2 by sol gel condensation. It happened with the help of external groups of glutamate and aspartate on the external surface of the virus [1,13,30,31]. Viral scaffolds can template the nucleation and assembly of inorganic materials. For example, cowpea chorotic mottle virus and cowpea mosaic virus have been used as nucleation cages for the mineralization of inorganic materials [43]. 6. CONCLUSIONS AND FUTURE PERSPECTIVES Microbial synthesis of nanoparticles has been emerged as an important branch of nanobiotechnology. Due to their rich diversity, microbes have the innate potential for the synthesis of nanoparticles and they could be regarded as potential biofactories for nanoparticles synthesis. Addition to microorganisms, their component can use for nanoparticle synthesis. Some biological molecules like fatty acids, amino acids, are used as template in the growth of semiconductor nanocrystals. In particular, by changing the ratio of different fatty acids (chain lengths), shapes of CdSe, CdS and CdTe nanocrystals can be achieved. Biological materials like DNA [32], protein cages [33], biolipid cylinders, viroid capsules, S-layers and multicellular superstructures have been used in template-mediated synthesis of inorganic nanoparticles. Although to improve synthesis rate and monodispersity of nanoparticles, factors such as microbial cultivation methods and downstream processes techniques should be heal [41] and combination of methods may be used such as photobiological methods [42], describe specific genes and characteristization of enzymes involved in the biosynthesis of nanoparticles is also required. Therefore, a complete knowledge of the molecular mechanisms involved in the microbial synthesis of nanoparticles is neseccery to control the size, shape and crystallinity of nanoparticles [43]. REFERENCES [1] K. B.Narayanan, N. Sakthivel. Biological synthesis of metal nanoparticles by microbes. Advances in Colloid and Interface Science 156 (2010) [2] T. Riddina, M. Gerickeb, C.G. Whiteleya, Biological synthesis of platinum nanoparticles: Effect of initial metal concentration. EMT J 46 (2010) [3] Kamyar Mollazadeh Moghaddam. An Introduction to Microbial Metal Nanoparticle Preparation Method. The journal of young investigations. Volume 19, Issue 19 January [4] P.Rajendran. Nanotechnology for Bioremediation of Metals. Nanotechnology for Bioremediation 2007, [5] Karbasian M, Atyabi SM, Siadat SD. Optimizing Nano-silver Formation by Fusarium oxysporum. American Journal of Agricultural and Biological Science 3(1): , [6] Deendayal Mandal. Mark E. Bolander.Debabrata Mukhopadhyay. The use of microorganisms for the formation of metal nanoparticles and their application. Appl Microbiol Biotechnol (2006) 69:

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