Protease constitutes a large and complex group of enzyme which plays an important nutritional and regulatory role in nature. Among five strains of endosulfan degrading bacterial isolates from rhizospheric soil samples of Crop Research Centre, GBPUAT, Pantnagar, strain EN-4 was found as the best producer of extracellular alkaline protease and selected for present study. The bacterium was identified as Bacillus subtilis strain EN4 on the basis of 16S rDNA gene sequencing. This protease had optimum activity at pH 10 and temperature 50°C. The enzyme was stable at alkaline pH range and upto a temperature of 70°C. The Km and Vmax values of EN4 protease were 0.55M and 22.2 U/ml, respectively. Partially purified alkaline protease was immobilized on calcium alginate beads with 59% activity recovery. The properties of immobilized alkaline protease were determined. The optimum pH and temperature for immobilized alkaline protease were 9 and 60°C, respectively.
Bacillus subtilis, Alkaline protease, Immobilization, Characterization
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