Abstract

Recent year, bacterial laccases are increasing interest in the field of industry and environmental applications especially decolorization of azo dyes. In industry, the dyes are present in stable nature including chemicals and lights. Due to these defects, the novel approaches are needed to removal of dyes before discharging into the environment. Among the various technologies, biological treatment methods and their strategies are very important, because of the decolorization and detoxification. Consecutively, biological mediated dyes removal are emerged with high potential especially microbes. Microbial laccases creates up new opportunities for their commercial applications. In this study, laccases were produced from Bacillus cereus (B. Cereus) and Pseudomonas parafulva (P. parafulva) by sub merged fermentation. For immobilization, the produced laccases were subjected to purify using 80% saturated ammonium sulphate and followed by dialysis. Then, crude laccases were immobilized through copper-alginate entrapment method. The maximum immobilized enzyme activity of the immobilized laccases were shown pH 8 at 50 °C and pH 7 at 40 °C for B. Cereus and P. parafulva respectively. In contrast, the normal enzyme activity was pH 10 at 40 °C and pH 8 at 40 °C were indicated for Bacillus cereus and P. parafulva respectively. Next, the free and immobilized laccases were performed the decolorization of three azo dyes T-blue, yellow GR and orange 3R, and exhibited that the 91.69 and 89.21% of Orange 3R were completely decolorized by both the B. Cereus and P. parafulva laccases when compared with free laccases enzymes. The confirmation of decolorization was monitored by UV–vis spectroscopy and FTIR spectroscopy, which clearly confirm the changes of peaks when compared with normal laccases. Finally, we have concluded that the B. Cereus and P. parafulva laccases are very important in azo dye decolorization and these used in future biological treatment of dyeing effluents.

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