Optimization studies of Protease enzyme in in-vitro conditions from Bacillus licheniformis

Protease enzymes are of immense commercial value and find its applications in various industrial sectors. Microbes serve as a major source for proteases enzyme especially Bacillus strains. In this study protease producing bacteria was isolated from the soil sample. Screening was done on skim milk casein agar media. The colony with the highest protease production was used as the test organisms and maintained on the nutrient agar slants. The isolated protease producing microorganism was characterized morphologically and biochemically by Bergey’s Manual of Determinative Bacteriology and identified to be as Bacillus licheniformis. Optimization studies were taken up with two physical and two chemical parameters. The organism showed maximum enzyme production at pH of 8.0, temperature 37°C, with 1.5% carbon source and 1.0% nitrogen source. Mass production was carried out for the protease enzyme with optimized media parameters and purified by centrifugation, ammonium sulphate precipitation (salting out), dialysis and Sephadex G-100 gel chromatography. The enzyme activity of the purified protease was assayed to be 6.66 U/ml. The 12% SDS-PAGE has revealed a unique linear band with a molecular weight of 52-53kDa. The results of this study revealed that the bacterial strain Bacillus licheniformis is a potent source for protease enzyme.