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In this study laccase mediated decolourization of Amido black, an azo dye was attempted. Laccase was extracted from the solid state fermentation of the white rot fungi Rigidoporus ulmarius on rice bran (50%). The enzyme activity was assayed using guaiacol as substrate and was found to be 0.0145 U/ml. The crude enzyme extract was employed for decolourization experiments. Optimization of the effect of contributing variables; pH, temperature, dye concentration and enzyme volume on decolourization was primarily analysed using OFAT (One Factor At a Time) approach. The optimum operating conditions for maximum decolourization were further statistically optimized by applying the Response surface methodology using Box-Behnken approach. A four factor three level Box-Behnken design generating 29 experimental runs was performed. A quadratic model was fitted and was analyzed using ANOVA at the 95% confidence interval. The determination coefficient R 2 was found to be 0.9616 indicating the significant fit of the model. The optimum operating conditions predicted by the model was found to be at constant incubation time of 24hrs, pH of 4; temperature of 30°C; dye concentration of 150mg/l and enzyme volume of 1.25ml producing 83.39% decolourization which was in good agreement with that of the predicted value, 83.24%.

Optimization of Enzymatic Decolourization of Amido black by Laccase from Rigidoporus ulmarius using Response Surface Methodology