Characterization and Purification of Protease enzyme

Protein is one of the three major food groups needed for proper nutrition. Proteolytic enzymes or proteinases are the group of enzymes whose catalytic function is to hydrolyze (breakdown) proteins. Production and partial purification of protease enzyme by Bacillus subtilis was the aim of this study. Bacillus subtilis was allowed to grow in shake flask broth culture, 3.5L and 7L fermenters for purpose of inducing protease enzyme. We are finding out the effect of minerals which are useful for the production of protease by Plackett Burman design. Minerals such as magnesium sulphate, potassium dihydrogen phosphate and manganese sulphate were showing the results for the production of protease. The protease enzyme was purified by ultra filtration, ammonium sulphate precipitation, dialysis, and lyophilization.The activity of protease was increased as there was increase in the enzyme concentration. Purified protease enzyme had a maximum activity at pH 9.0 of carbonate buffer and the optimum incubation time was 48hr. The protease assay is done for the crude enzyme at different temperature. It showed greater activity at 50°C but after that it started decreasing the activity so, we had selected the temperature at 40°C for the good activity.