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Extra Cellular Bio Synthesis of Silver Nano Particles Using the Fungus Fusarium Semitectum

Extra Cellular Bio Synthesis of Silver Nano Particles Using the Fungus Fusarium Semitectum

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02/04/2011

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Extracellular biosynthesis of silver nanoparticles using thefungus
Fusarium semitectum
S. Basavaraja
a,b
, S.D. Balaji
a,b
, ArunkumarLagashetty
c
, A.H. Rajasab
d
,A. Venkataraman
a,b,
*
a
 Department of Materials Science, Gulbarga University, Gulbarga 585106, Karnataka, India
b
 Department of Chemistry, Gulbarga University, Gulbarga 585106, Karnataka, India
c
 Appa Institute of Engineering and Technology, Gulbarga 585102, Karnataka, India
d
 Department of Botany, Gulbarga University, Gulbarga 585106, Karnataka, India
Received 27 January 2007; received in revised form 14 April 2007; accepted 3 June 2007Available online 12 June 2007
Abstract
Development of environmental friendly procedures for the synthesis of metal nanoparticles through biological processes isevolving into an important branch of nanobiotechnology. In this paper, we report on the use of fungus
Fusarium semitectum
’’ forthe extracellular synthesis of silver nanoparticles from silver nitrate solution (i.e. through the reduction of Ag
+
to Ag
0
). Highlystable and crystalline silver nanoparticles are produced in solution by treating the filtrate of the fungus
F. semitectum
with theaqueous silver nitrate solution. The formations of nanoparticles are understood from the UV–vis and X-ray diffraction studies.Transmissionelectronmicroscopyofthesilverparticlesindicatedthattheyrangedinsizefrom10to60 nmandaremostlysphericalin shape. Interestingly the colloidal suspensions of silver nanoparticles are stable for many weeks. Possible medicinal applicationsof these silver nanoparticles are envisaged.
#
2007 Elsevier Ltd. All rights reserved.
Keywords:
A. Metals; C. X-ray diffraction; C. Infrared spectroscopy; D. Microstructure
1. Introduction
The nanoparticles of noble metals are found to have potential applications in various fields like microelectronics[1], optical devices[2], catalysis[3]and drug delivery system[4], etc. The noble metal nanoparticles exhibit new physico-chemical properties which are not observed either in the individual molecules, or in the bulk metals[3,5,6].For example, gold and silver nanoparticles exhibit strong absorption of electromagnetic waves in thevisible range dueto surface plasmon resonance (SPR). SPR is caused due to collective oscillations of the conduction electrons of nanoparticles upon irradiation with visible light[7]. The SPR is highly influenced by shape and size of thenanoparticles. Recently, the absorption spectra of individual silver nanoparticles were correlated with their size andshape determined by transmission electron microscopy (TEM)[8]. The results indicate that spherical and roughlyspherical nanoparticles absorb in the blue region of the spectrum, while decahedral nanoparticles and particles with
www.elsevier.com/locate/matresbuMaterials Research Bulletin 43 (2008) 1164–1170* Corresponding author at: Department of Materials Science, Gulbarga University, Gulbarga 585106, Karnataka, India. Tel.: +91 8472 263295;fax: +91 8472 263206.
E-mail address:
raman_chem@rediffmail.com(A. Venkataraman).0025-5408/$ see front matter
#
2007 Elsevier Ltd. All rights reserved.doi:10.1016/j.materresbull.2007.06.020
 
triangular cross-sections absorb in the green and red part of the spectrum, respectively. The width and position of theSPR not only depends on the particle size as suggested earlier, but also on the chemical properties of thenanocrystalline surface, referred to as chemical interface damping[9]. Thus, an exquisite control of size, composition,morphology, stability and environmental friendly synthesis are the features which are highly desirable. Though thereare several physical and chemical methods for synthesis of metallic nanoparticles, to achieve these objectives,researchers turned to biological systems. Many organisms, both unicellular and multicellular are known to produceinorganic nanomaterials either intracellularly[10]or extracellularly[10], even though the actual mechanisms are not fully understood because of the complexity of most biological reactions. Varieties of inorganic nanomaterials aresynthesised by biological processes using bacteria, yeast and fungi[11–13]. While intracellular synthesis in principlemay accomplish a better control over the size and shape distributions of the nanoparticles, product harvesting, andrecovery are more cumbersome and expensive. The extracellular synthesis by comparison is more adaptable to thesynthesis of a wider range of nanoparticles systems. Among various metal nanoparticles, silver nanoparticles haveseveral important applications in the field of biolabelling[14], sensors, antimicrobial agents and filters[15]and hence are being intensively studied employing
Fusarium oxysporum
Pseudomonas stutzeri
Rhodococcus
sp.[16],
Thermomonospora
Phaenero chaete chrysosporium
[17], etc. In the present investigation we report theextracellular biosynthesis of silver nanoparticles employing the fungus
Fusarium semitectum
.
F. semitectum
iscommonly available fungus found in marshland regions. In literature we have not come across using this fungus forformation and stabilization of silver nanoparticles in aqueous system. The local environment suits for this fungus, andalso,thisfungusisrelatedto
F.oxysporum
inmanyaspects.Hencewehaveundertakenthisfungusinthepresentstudy.The present study includes time dependent formation of silver nanoparticles employing UV–vis spectrophotometer,size and morphology by employing TEM, structure from powder X-ray diffraction (XRD) technique andunderstanding of protein–silver nanoparticles interaction from Fourier transform infrared (FT-IR) spectroscopy.
2. Experimental
The fungus
F. semitectum
was obtained from Agharkar Research Institute, Pune, India and maintained on potatodextrose agar plants. To prepare biomass for biosynthesis studies the fungal was grown aerobically in a liquid mediacontaining (g/l) KH
2
PO
4
, 7.0; K 
2
HPO
4
, 2.0; MgSO
4
Á
7H
2
O, 0.1; (NH
2
)SO
4
, 1.0; yeast extract, 0.6; and glucose, 10.0.The flasks were inoculated and then incubated on orbital shaker at 27
8
C and agitated at 150 rpm. The biomass washarvested after 72 h of growth by sieving through a plastic sieve,followed by extensivewashing with distilled water toremove any medium component from the biomass. Typically 20 g (wet weight) was brought in contact with 100 ml of double distilled water for 72 h at 27
8
C in an Erlenmeyer flask and agitated in the same conditions as described earlier.After the incubation, the cell filtrate was obtained by passing it through Whatman filter paper no. 1. The resultantfiltrate was then mixed with 100 ml of carefully weighed 10
À
3
M AgNO
3
solution in a 250 ml Erlenmeyer flask andkept on a shaker at 27
8
C. Periodically aliquots of the reaction solution were removed and subjected to UV–visspectroscopy measurements. The UV–vis spectroscopy measurements wereperformed on an Elico spectrophotometerat a resolution of 1 nm from 200 to 800 nm. Films of silver nanoparticles were formed on Si(1 1 1) substrates by drop-coating the colloidal nanoparticles for XRD study. The data was obtained with a Siemens X-ray diffractometer(Japan), and the target was Cu K 
a
(
l
= 1.54 A˚). The generator was operated as 30 kVand with a 20 mA current. Thescanning range (2
) was selected from 10
8
to 80
8
angles. A scanning speed of 1
8
 /min and a chart speed of 20 mm/minwere used for the precise determination of the lattice parameters. High-purity silicon powder was used as an internalstandard. The coherently diffracting crystallographic domain size (
XRD
) of the silver nanoparticles was calculatedfrom X-ray diffraction (XRD) line broadening after subtracting the contribution from the Cu
a
component(Rachinger correction) and correcting for the instrumental width. The integral line width was used in the Scherrerformula to calculate
XRD
of the (1 1 1) plane.Fortransmission electron microscopy (TEM),the sampleswere prepared byadropofcolloidal solution ofsilveronacarboncoated coppergridandsettingthe dropdrycompletelyinavacuumdesiccator.TheTEM image ofthe samplewas obtained using Technai-20 Philips transmission electron microscope. The transmission electron microscope wasoperated at 190 keV.The sediment particles obtained after 2 months are taken for the FT-IR studies. The FT-IR spectrum of the samplewas recorded on a Perkin-Elmer FT-IR Spectrum ONE in the range 4000–400 cm
À
1
at a resolution of 4 cm
À
1
bymaking the KBr pellet with the silver nanoparticles.
S. Basavaraja et al./Materials Research Bulletin 43 (2008) 1164–1170
1165
 
3. Results and discussion
The detailed study on extracellular biosynthesis of silver nanoparticles by
F. semitectum
was carried out and isreported in this work.Fig. 1shows conical flasks containing the filtrate of the
F. semitectum
biomass with Ag
+
ions atthe beginning and after 2 days of the reaction, respectively. It is observed that the colour of the solution turned fromcolourless to brown after 24 h of the reaction, indicating the formation of silver nanoparticles. It is well known thatsilver nanoparticles exhibit yellowish brown colour in water; this colour arises due to excitation of surface plasmonvibrations inthemetal nanoparticles[18].This important observationindicates thatthe reduction ofthe Ag
+
ions takesplace extracellularly. The formation and stability of the reduced silver nanoparticles in the colloidal solution wasmonitored by using UV–vis spectral analysis.An UV–vis spectrum is one of the important techniques to ascertain the formation of metal nanoparticle, providedsurface plasmon resonance exists for the metal. The UV–vis spectra recorded from
F. semitectum
reaction vessels atdifferent time intervals of reaction are plotted and is shown inFig. 2. The time at which the aliquots were removed for
S. Basavaraja et al./Materials Research Bulletin 43 (2008) 1164–1170
1166Fig.1. Pictureofconicalflaskscontainingthefiltrateofthe
Fusariumsemitectum
biomassinaqueoussolutionof10
À
3
MAgNO
3
atthebeginningof the reaction (flask 1) and after 2 days of reaction (flask 2).Fig. 2. UV–vis spectra recorded as a function of time of reaction of an aqueous solution of 10
À
3
M AgNO
3
with the filtrate of the fungal biomass.The time of reaction is indicated next to the respective curves.

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