PROTEASE PRODUCTION FROM Bacillus safensis IN SUBMERGED FERMENTATION USING RESPONSE SURFACE METHODOLOGY

In present study, various culture parameters for Bacillus safensis were optimized in submerged fermentation for alkaline protease production using response surface methodology. Nutritional components were screened through Placket-Burman design. The optimized culture parameters were, 1% inoculum size, initial medium of pH 9.0, incubation temperature of 30 degrees C and fermentation time of 72 h. Among nutritional parameters, casein, KH2PO4 and peptone were found significant which was further optimized through Box-Bhenken Design with three levels. The experimental results showed that 4.0% casein, 0.05% KH2PO4 and 0.575% peptone yielded highest protease production. The proposed model was significant which clearly explained the response. Partial characterization revealed that crude protease produced by Bacillus safensis showed optimum activity at pH 8.0 and at 40 degrees C. Kinetics revealed the K-m and V-max values of 1.027 mg/ml and 67.57 mg/ml/min using casein as substrate, respectively.