Enhancement of Alkaline Protease Production by Bacillus Species Through Random Mutagenesis

Bacillus species is a potent strain which produced the alkaline protease and it is further mutated to yield more amount of enzyme. An alkaline protease producing strain of Bacillus species was isolated from soil. The isolates were purified and screened for the production of protease enzyme. The conditions for enzyme production were optimized under solid substrate fermentation (SSF) and submerged fermentation (SF). Maximum enzyme production occurred at 50°C, pH 8. 0 at 72 hours of incubation under SF using Reese medium. Among various combinations of natural substrates used for enzyme production by SSF, mixture of paddy husk, ground nut shell 3:2 ratio served as a suitable and best for alkaline protease production. It was thermo stable and retained full activity after 1hour of incubation at 50°C. Enzyme was purified by gel filtration, Iron exchange chromatography and molecular weight of 31 KDa was confirmed by SDS-PAGE. The enzyme was retained with a maximum and minimum activity of 84. 9% at 10 minutes and 1% at 40 minutes and completely lost at 50 minutes in the presence of detergents such as Surf, RIN, Ariel, Wheel, indicating its suitability for application in detergent industry. The enzyme activity was extended with the help of immobilization technique using sodium alginate and polyacrylamide gel.