PURIFICATION AND CHARACTERIZATION OF PROTEASE ENZYME FROM NATIVE ISOLATE BACILLUS SUBTILIS AND ITS COMPATABILITY WITH COMMERCIAL DETERGENTS

The present study describes the purification characterization of crude protease from Bacillus subtilis and its evaluation in detergent. An alkaline protease producing B.subtilis was isolated from local soil samples were collected from Local Garden. The bacterial culture B.subtilis showed luxurious growth in Nutrient agar at pH (7.2) and at a temperature 37°C. The bacterium is mesophilic in nature. The enzyme was purified in a 2-step procedure involving ammonium sulfate precipitation and Sephadex G-200 gel permeation chromatography. The molecular weight of the enzyme determined by SDS-PAGE was found to be(30KDa) and was purified (112)fold with a yield of (6%). The highest protease activity was found to be atpH (10.0) using glycine-NaOH buffer. The enzyme was almost 100% stable at60°C even after (5) hours of incubation.Most of the metal ions tested had a stimulatory effect (Ca2+, Mg2+, and Mn2+) or a slight inhibitory effect (other ions) on enzyme activity. The protease showed excellent stability and compatibility in the presence of locally available detergents. The enzyme retained more than 50% activity with most of the detergents tested even after 2 hours incubation at 60°C after the supplementation of(5mMCaCl2)and(1M glycine).The purpose of the research was to study designed to isolate purificationand characterize the protease enzyme from Bacillus subtilis and its compatibility with commercial detergents.