Purification, Partial Characterization and Applications of Alkaline Protease from Bacillus subtilis NCIM No. 2724

Thermostable alkaline protease production from Bacillus subtilis NCIM No. 2724, its purification and characterization was investigated. The purified enzyme revealed MW of 26 KDa. This enzyme is stable at a pH and temperature range of 7–11 with optimum pH at 10.0 and 30 to 70°C with optimum activity at 60°C, respectively. The enzyme is strongly activated by metal ions such as Ca²⁺, Mg²⁺ and Mn²⁺ while the activity is negatively regulated by EDTA, urea, guanidine hydrochloride and SDS. Whereas >50% activity of the purified enzyme was observed in the presence of commercial detergents tested, at 60°C. Blood stain removal was effective with this enzyme when supplemented with CaCl₂ and glycine.