geometrical symmetry. Even though many biotechnolog-ical applications such as the remediation of toxic metals employmicroorganisms such as bacteria and fungi, such microorgan-isms are recently found as possible eco-friendly nanofactories. This study demonstrated the extracellular synthesis of stable silver nanoparticles using the fungus,
The fungus culture,
(NCIM 650) wasobtained from National Chemical Laboratory culture collection,Pune, India. All chemicals used were of analytical grade.
was grown in yeast malt broth containing dextrose, 10 g/ L; peptic digest of animal tissue, 5 g/L; yeast extract, 3 g/L; andmalt extract, 3 g/L. The final pH was adjusted to 6.2±0.2. Theflasks were incubated in the environmental shaker at 200 rpm at 37 °C. After 5 days of incubation, the mycelium was separated by filtration and washed thrice with Milli-Q deionized water.The washed mycelium (5 g fresh weight) was challenged with100 mL of 1 mM silver nitrate solution (prepared in deionizedwater) and incubated in shaker at 200 rpm in dark condition at 37 °C. Simultaneously, a positive control of incubating thefungus mycelium with deionized water and a negative controlcontaining only silver nitrate solution were maintained under same conditions.
visible spectral analysis
Change in colour was observed in the silver nitrate solu-tion incubated with the
. The UV
visible spectra of this solution was recorded in Specord 50 ANALYTIKJENA®Spectrophotometer, from 200 to 800 nm, at an interval of 24 h.The silver nanoparticles dispersed in water by ultrasonicationwas kept at room temperature (37 °C) for 3 months and theabsorption at 420 nm was measured continuously, to determinetheir stability.
2.3. X-ray diffraction analysis
The fungal mycelium embedded with the silver nanoparticleswas freeze-dried, powdered and used for XRD analysis. Thespectra were recorded in Philips® automatic X-ray Diffractom-eter with Philips® PW 1830 X-ray generator. The diffractedintensities were recorded from 30° to 80° 2
angles.Simultaneously, the control fungal sample was also analyzedas per the above-mentioned procedure. For standard, the silver metal powder was subjected to XRD analysis.
2.4. SEM, TEM and electron diffraction analysis
The freeze-dried mycelial mats (positive control and silver nitrate treated sample) were mounted on specimen stubs withdouble-sided adhesive tape and coated with gold/palladium in asputter coater and examined under Philips® XL 30 SEM at 12
15 kV with a tilt angle of 45°. For TEM, a drop of aqueoussolution containing the silver nanomaterials was placed on thecarbon coated copper grids and dried under Infrared lamp.
Fig. 1. UV
visible spectra of fungal filtrate as a function of time. Curve Acorresponds to positive control, while B, C, D and E correspond to that of incubations with silver nitrate (1 mM) after 24 h, 48 h, 72 h and 96 h,respectively. The peak at 420 nm corresponds to the surface plasmon resonanceof silver nanoparticles. The inset is a plot of the absorbance at 420 nm as afunction of time, corresponding to the spectra shown in the main figure.Fig. 2. X-ray diffraction pattern of the dried fungal mycelium with and without the impregnated silver nanoparticles. In spectrum
(recorded for pure metallicsilver powder), the diffractions at 38.5°, 44° and 64.5° 2
can be indexed to the(111), (200) and (220) planes of the face-centered cubic (fcc) silver, respectively.Spectrum
corresponds to the fungal mycelial mat without silver nitratetreatment. Spectrum
corresponds to the fungal mycelial mat after beingtreated with silver nitrate. Here, the filled circles (
) indicate the peakscorresponding to that of fungal mycelial mat (semi-crystalline chitinmicro fibrils), stars (
) indicate the peaks produced by metallic silver, whiletwo new peaks (?) are formed due to the interaction of silver with the fungalcell wall.1414
N. Vigneshwaran et al. / Materials Letters 61 (2007) 1413