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Decolorization of the dye congo red by Aspergillus niger silver nanoparticle R.Nithya1 and R.Ragunathan*,

Dean, Faculty of Science and 1PhD Research Scholar,

Department of Biotechnology, Shri Nehru Maha Vidhyalaya College of Arts and Science, Coimbatore-21, Tamilnadu. Corresponding author: Abstract Bioremediation using a variety of microbes for the degradation of xenobiotics seems a green solution to the problem of environmental pollution. Microbes have been gifted by nature with the ability of degrading a wide spectrum of environmental pollutants. Different fungi have the potentials to degrade complex and recalcitrant organic compounds into simpler fragments; sometimes achieving complete mineralization. Nanomaterials reveal good result than other techniques used in waste water treatment because of its high surface area. It is suggested that these may be used in future at large scale water purification. In the present study we have reported the decolorization of the dye congored by using Aspergillus niger silver nanoparticle and its comparison with plain culture. The silver nanoparticle was synthesized at 1mM concentration. Detailed characterization of nanoparticle using UV-Visible studies and Scanning electron microscopic studies confirmed the presence of 200nm sized particles. These particles were then checked for their efficiency to decolorize the dye congored. The nanoparticle efficiently decolorized the dye within 24hour of incubation where as the plain culture (Control) decolorization continued upto 48 hours. In our experiment we found that Aspergillus niger silver nanoparticle as an efficient decolorizer against the dye congored. Key words: Silver nanoparticle, decolorization, azo dye, waste water. Introduction Removal of dyes from textile waste effluents has been carried out by physical, chemical and biological methods, such as flocculation, membrane filtration, electrochemical techniques, ozonation, coagulation, adsorption and fungal discoloration (Fu and Tiraraghavan, 2004). Fungal bioremediation is becoming an attractive option for removal of dyes from industrial effluents as microorganisms are natures tools for cleaning the environment. Without fungi and their decomposing activities, the world would have become a heap of dead bodies of plants and animals. Removal of dyes from industrial waste waters is of global concern because dyes cause many problems in aqueous environments. Dyes may significantly affect photosynthetic activity

in aquatic life because of reduced light penetration and may also be toxic to some aquatic life due to the presence of aromatics, metals, chlorides, etc. (Daneshvar et al., 2007). Nanotechnology enables the development of nanoscale particles of metals with novel and distinctive physic-chemical properties, and a broad range of scientific and technological applications (Moore 2006). Another potential use of silver nanoparticles in water filters in wastewater treatment plants. At nanoscale silver exhibits remarkably unusual physical, chemical and biological properties (Evan off and Chumanov 2005; Chen and Schluesener 2007). Recently it was shown that silver ions may be reduced extracellularly using fungus Phanerochaete chrysoporium (Vigneshwaran et al., 2007) and Pleurotus sajor caju (Nithya and Ragunathan 2009). In this paper we have made an attempt to decolorize the dye congored by silver nanoparticle synthesized by using Aspergillus nigerMTCC 1784 and its comparison with its plain culture. Materials and Methods: Organism: The organism used in this study was Aspergillus nigerMTCC 1784 and it was maintained on 2% malt agar plates. Synthesis of silver nanoparticle The organism was allowed to grow in the biomass production broth containing Glucose 10g/l, Potassium dihydrogen orthophosphate 1g/l, potassium chloride 0.5g/l, Magnesium sulphate 5g/l, Ferrous sulphate 0.1g/l, Sodium nitrate 2g/l, Malt extract 1g/l, yeast extract 1g/l at a pH of 6.0. and after 7 days the biomass was separated and allowed to grow in deionised water for 3days.To the filtered biomass 1mM final concentration AgNo3was added and incubated in dark conditions at 30C under shaking conditions and the formation of nanoparticle was examined under UVvisible spectrophotometer at 24hr time interval. Characterization of the nanoparticle The particle was characterized by UV-visible studies and the particles were subjected to SEM studies for their size determination. Decolorization studies For decolorization study, 250 mL Erlenmeyer flasks containing 125 mL solutions of congo red was prepared in the media containing 30 g sucrose, 3 g NaNO3, 0.5 g KCl, 0.5 g MgSO4.7H2O, 0.01g FeSO4.7H2O, 1 g K2HPO4 and 15 g agar per liter was used. Final pH of the medium is 7.3 0.2. The Aspergillus niger silver nanoparticle was added to the above media which is indicated as test. Similarly plain culture (Aspergillus niger) was also added to this media separately which serves as control for the above. Congo red of 50M concentration was used in this study. The flasks were incubated at 30C under shaking conditions. After 24hr interval

samples were withdrawn, filtered and centrifuged at 4400rpm for 5mins and the supernatants was analyzed spectrophotometrically using UV-Visible spectrophotometer at 498nm. Results The Aspergillus niger silver nanoparticle was formed after 48hours of the incubation with 1mM silver nitrate. Characterization studies The UV-Visible studies indicated the surface Plasmon resonance at 379nm which depicts the formation of silver nanoparticle. (Fig.1) SEM studies The SEM micrographs of nanoparticle obtained in the filtrate showed that silver nanoparticles are spherical shaped ,well distributed without aggregation and an average size of about 200nm. (Fig.2) Decolorization studies A significant decolorization rate was observed for the dye Congo red. The Aspergillus niger silver nanoparticle effectively decolorized 85.8% of dye within 24 hour incubation and the dye was fully decolorized within 48 hour of incubation. Whereas the plain culture (Aspergillus niger) was able to degrade only 76% of dye at the same incubation conditions and complete decolorization was observed after 48 hour incubation.(Fig.3A and 3B),Graph.1

Fig.1 UV-Visible graph of the nanoparticle

Fig.2 SEM image of the Aspergillus niger AgNP

Fig.3A Decolorization of Aspergillus niger(Plain culture)

Fig.3BDecolrizationof Aspergillus nigersilver nanoparticle

Graph.1. % of decolorizationof the dye congo red by Aspergillusniger silver nanoparticle and Aspergillus niger (plain culture) Discussion (Hazrat Ali et al.,2009) has reported 96.91% of decolorization of MG by Aspergillus flavus In this study when compared with nanoparticle the plain culture could degrade only 76% of the dye this might me due to the complex nature of the dye. A slower rate of decolorization was attributed to higher molecular weight, structural complexicity and the presence of inhibitory groups like NO2 and SO3Na in the dyes (Hu and Wu, 2001). After decoloration, the absorbance at wavelength 498 nm decreased with the largest decrease at the peak. A general decrease of this kind is usually attributed to dye degradation rather than adsorption (Wesenberg et al., 2002). These data show that decolorization was caused by degradation of dye. (Sadowski et al .,2008 and Kalishwaralal et al .,2008) reported the nanoparticles synthesized in the size range of 100nm and 50nm and in this paper we obtain particles in the size range of 200nmThis preliminary study shows silver nanoparticle can be effectively used for the decolorization processes. Conclusion The present study revealed the ability of the Aspergillus niger silver nanoparticle to decolorize congored. From the results we conclude that the nanoparticle decolorize better than the plain culture of the same strain. These preliminary results suggest that silver nanoparticles can be used for treatment of textile effluents. The development of such particles may be considered a breakthrough in the field for the efficient clean up of the dyes on large scale process since they are easy to synthesize on large scale and cost effective. Further research on the role of pH, temperature and other parameters will be carried out.

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