Asian Journal of Microbiology, Biotechnology & Environmental Sciences Paper

ISOLATION, PURIFICATION AND CHARACTERIZATION OF PROTEASE ENZYME FROM BACILLUS SP., POSSIBLE USE IN BIODIESEL PRODUCTION

A. ANITHA AND M. REBEETH

Abstract

The identified genus Bacillus sp (C 1) producing maximum protease enzyme was isolated from the slaughterhouse soil collected from Othakkalmandapam, Coimbatore, Tamil Nadu, India. Mass culture was done in suitable growth media to extract the enzyme protease. The enzyme was purified by 3-step procedure involving ammonium sulphate precipitation, dialysis and sephadex G-200 gel filtration chromatography. The specific activity of the purified enzyme was found to be 1000 IU/mL. The protein of 0.4 mg/mL and the purification fold of 3.91 were obtained and it produced a single band on SDS PAGE with molecular weight of 68 KDa. The maximum activity of the purified protease enzyme was found at the optimum pH of 11 and the optimum temperature of 600C with Vmax of 700mg and the Km of 130mg/ml. Thermostablitiy of the protease enzyme indicated that it is suitable for industrial purposes. A new alkaline protease having application in biodiesel production. Degumming is the first step in biodiesel production. It was carried out using phosphoric acid and the isolated protease enzyme. The physical properties showed the removal of phosphatides in crude oil by the treatment of enzyme is more compared with acid degummed oil. So the enzymatic degumming is considered as more sophisticated one in terms of specificity, minimal waste production, etc., than other degumming process.