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Green synthesis of protein capped silver nanoparticles from phytopathogenic fungus Macrophomina phaseolina (Tassi) Goid with antimicrobial properties against multidrug-resistant bacteria.

Chowdhury S, Basu A, Kundu S - Nanoscale Res Lett (2014)

Bottom Line: These nanoparticles were found to be naturally protein coated.Antimicrobial activities of the silver nanoparticles against human as well as plant pathogenic multidrug-resistant bacteria were assayed.The particles showed inhibitory effect on the growth kinetics of human and plant bacteria.

View Article: PubMed Central - HTML - PubMed

Affiliation: Molecular and Applied Mycology and Plant Pathology Laboratory, Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata 700019, India.

ABSTRACT
In recent years, green synthesis of nanoparticles, i.e., synthesizing nanoparticles using biological sources like bacteria, algae, fungus, or plant extracts have attracted much attention due to its environment-friendly and economic aspects. The present study demonstrates an eco-friendly and low-cost method of biosynthesis of silver nanoparticles using cell-free filtrate of phytopathogenic fungus Macrophomina phaseolina. UV-visible spectrum showed a peak at 450 nm corresponding to the plasmon absorbance of silver nanoparticles. Scanning electron microscopy (SEM), atomic force microscopy (AFM), and transmission electron microscopy (TEM) revealed the presence of spherical silver nanoparticles of the size range 5 to 40 nm, most of these being 16 to 20 nm in diameter. X-ray diffraction (XRD) spectrum of the nanoparticles exhibited 2θ values corresponding to silver nanoparticles. These nanoparticles were found to be naturally protein coated. SDS-PAGE analysis showed the presence of an 85-kDa protein band responsible for capping and stabilization of the silver nanoparticles. Antimicrobial activities of the silver nanoparticles against human as well as plant pathogenic multidrug-resistant bacteria were assayed. The particles showed inhibitory effect on the growth kinetics of human and plant bacteria. Furthermore, the genotoxic potential of the silver nanoparticles with increasing concentrations was evaluated by DNA fragmentation studies using plasmid DNA.

No MeSH data available.


Related in: MedlinePlus

Synthesis of silver nanoparticles using cell-free filtrate of Macrophomina phaseolina and spectroscopic analysis. (a) Photograph of 1 mM AgNO3 solution without cell filtrate (1, control), mycelia-free cell filtrate of M. phaseolina (2), and 1 mM AgNO3 with cell filtrate after 24-h incubation at 28°C (3). (b) UV–vis spectra recorded as a function of time of reaction at 24, 48, and 72 h of incubation of an aqueous solution of 1 mM AgNO3 with the M. phaseolina cell filtrate showing absorption peak at 450 nm.
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Figure 1: Synthesis of silver nanoparticles using cell-free filtrate of Macrophomina phaseolina and spectroscopic analysis. (a) Photograph of 1 mM AgNO3 solution without cell filtrate (1, control), mycelia-free cell filtrate of M. phaseolina (2), and 1 mM AgNO3 with cell filtrate after 24-h incubation at 28°C (3). (b) UV–vis spectra recorded as a function of time of reaction at 24, 48, and 72 h of incubation of an aqueous solution of 1 mM AgNO3 with the M. phaseolina cell filtrate showing absorption peak at 450 nm.

Mentions: The cell-free filtrate of M. phaseolina was used for the biosynthesis of the silver nanoparticles as described in methods. Figure 1a shows that AgNO3 solution itself is colorless (tube 1). The fungal cell filtrate incubated without AgNO3 (tube 2) was also almost colorless. The fungal cell filtrate, after incubation with 1 mM AgNO3 (tube 3), underwent a distinct change in its color to brown within 24 h, which indicated the formation of silver nanoparticles due to the conversion of Ag+ ions to elemental Ag by extracellular reductase activity of M. phaseolina filtrate. The color intensity of the silver nanoparticle solution persisted even after 72 h, which indicated that the particles were well dispersed and stable in the solution. The mycosynthesis of silver nanoparticles involves trapping of Ag + ions at the surface of the fungal cells and the subsequent reduction of the silver ions by the extracellular enzymes like naphthoquinones and anthraquinones present in the fungal system. One earlier study with Fusarium oxysporum shows that NADPH-dependent nitrate reductase and shuttle quinine extracellular process are responsible for nanoparticle formation [31]. Extracellular secretion of enzymes is especially advantageous for large-scale nanoparticle synthesis since large quantities of relatively pure enzyme can be obtained, free from other cellular proteins associated with the organism. The nanoparticles thus produced can be easily isolated by filtering from the reaction mix [28].


Green synthesis of protein capped silver nanoparticles from phytopathogenic fungus Macrophomina phaseolina (Tassi) Goid with antimicrobial properties against multidrug-resistant bacteria.

Chowdhury S, Basu A, Kundu S - Nanoscale Res Lett (2014)

Synthesis of silver nanoparticles using cell-free filtrate of Macrophomina phaseolina and spectroscopic analysis. (a) Photograph of 1 mM AgNO3 solution without cell filtrate (1, control), mycelia-free cell filtrate of M. phaseolina (2), and 1 mM AgNO3 with cell filtrate after 24-h incubation at 28°C (3). (b) UV–vis spectra recorded as a function of time of reaction at 24, 48, and 72 h of incubation of an aqueous solution of 1 mM AgNO3 with the M. phaseolina cell filtrate showing absorption peak at 450 nm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4114801&req=5

Figure 1: Synthesis of silver nanoparticles using cell-free filtrate of Macrophomina phaseolina and spectroscopic analysis. (a) Photograph of 1 mM AgNO3 solution without cell filtrate (1, control), mycelia-free cell filtrate of M. phaseolina (2), and 1 mM AgNO3 with cell filtrate after 24-h incubation at 28°C (3). (b) UV–vis spectra recorded as a function of time of reaction at 24, 48, and 72 h of incubation of an aqueous solution of 1 mM AgNO3 with the M. phaseolina cell filtrate showing absorption peak at 450 nm.
Mentions: The cell-free filtrate of M. phaseolina was used for the biosynthesis of the silver nanoparticles as described in methods. Figure 1a shows that AgNO3 solution itself is colorless (tube 1). The fungal cell filtrate incubated without AgNO3 (tube 2) was also almost colorless. The fungal cell filtrate, after incubation with 1 mM AgNO3 (tube 3), underwent a distinct change in its color to brown within 24 h, which indicated the formation of silver nanoparticles due to the conversion of Ag+ ions to elemental Ag by extracellular reductase activity of M. phaseolina filtrate. The color intensity of the silver nanoparticle solution persisted even after 72 h, which indicated that the particles were well dispersed and stable in the solution. The mycosynthesis of silver nanoparticles involves trapping of Ag + ions at the surface of the fungal cells and the subsequent reduction of the silver ions by the extracellular enzymes like naphthoquinones and anthraquinones present in the fungal system. One earlier study with Fusarium oxysporum shows that NADPH-dependent nitrate reductase and shuttle quinine extracellular process are responsible for nanoparticle formation [31]. Extracellular secretion of enzymes is especially advantageous for large-scale nanoparticle synthesis since large quantities of relatively pure enzyme can be obtained, free from other cellular proteins associated with the organism. The nanoparticles thus produced can be easily isolated by filtering from the reaction mix [28].

Bottom Line: These nanoparticles were found to be naturally protein coated.Antimicrobial activities of the silver nanoparticles against human as well as plant pathogenic multidrug-resistant bacteria were assayed.The particles showed inhibitory effect on the growth kinetics of human and plant bacteria.

View Article: PubMed Central - HTML - PubMed

Affiliation: Molecular and Applied Mycology and Plant Pathology Laboratory, Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata 700019, India.

ABSTRACT
In recent years, green synthesis of nanoparticles, i.e., synthesizing nanoparticles using biological sources like bacteria, algae, fungus, or plant extracts have attracted much attention due to its environment-friendly and economic aspects. The present study demonstrates an eco-friendly and low-cost method of biosynthesis of silver nanoparticles using cell-free filtrate of phytopathogenic fungus Macrophomina phaseolina. UV-visible spectrum showed a peak at 450 nm corresponding to the plasmon absorbance of silver nanoparticles. Scanning electron microscopy (SEM), atomic force microscopy (AFM), and transmission electron microscopy (TEM) revealed the presence of spherical silver nanoparticles of the size range 5 to 40 nm, most of these being 16 to 20 nm in diameter. X-ray diffraction (XRD) spectrum of the nanoparticles exhibited 2θ values corresponding to silver nanoparticles. These nanoparticles were found to be naturally protein coated. SDS-PAGE analysis showed the presence of an 85-kDa protein band responsible for capping and stabilization of the silver nanoparticles. Antimicrobial activities of the silver nanoparticles against human as well as plant pathogenic multidrug-resistant bacteria were assayed. The particles showed inhibitory effect on the growth kinetics of human and plant bacteria. Furthermore, the genotoxic potential of the silver nanoparticles with increasing concentrations was evaluated by DNA fragmentation studies using plasmid DNA.

No MeSH data available.


Related in: MedlinePlus