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.
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