An alkalophilic and thermostable protease producing Bacillus licheniformis strain BIHPUR 0104 was isolated from poultry farm. Response surface methodology by Box-Behenken design was employed for the enhanced production of protease using independent variables selected through one variable at one time approach. Under the optimized condition of sucrose (1.5% w/v), temperature (40°C) and incubation period (24 h), the protease production from strain BIHPUR 0104 was enhanced 2 fold compared to one variable at one time approach. The protease experimental yield (483.23 U/ml) closely matched the predicted value (480.74 U/ml) by the statistical model (R2=0.9997). The p values showed that the independent variables had highly significant effect on proteolytic activity (p<0.0001) of the strain tested. The crude protease obtained from strain BIHPUR 0104 showed stability at a wide range of pH (9) and temperature (50°C) indicating thermo-alkali stable nature of the crude enzyme.
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