PRODUCTION, PURIFICATION AND CHARACTERIZATION OF BACILLUS LIPASE

The lipolytic activities in the supernatant fractions of Bacillus cere us and Bacillus coagulans cultures were investigated. Aeration, agitat ion, different media, emulsified oils, inoculum size and phase of grow th affected lipase production. Aeration was essential for lipase produ ction (air:medium ratio 4:1) and produced the highest activity. The li polytic activity reached a maximum level after incubation for two days with continuous agitation. It was also elevated by the presence of ei ther olive oil or tributyrin and with lesser extent in the presence of castor oil. The enzyme levels were drastically reduced in the presenc e of animal fat, cotton seed oil, margarine or glycerol. The extracell ular lipase enzyme from Bacillus cereus was purified with 46.2% overal l recovery throught too steps, an acetone precipitation of the whole s upernatant and purification by gel filtration on sephadex G-100. The e fficiency of the purification process was evaluated through the polyac rylamide gel electrophoresis. The enzyme has an optimum pH 7.5 at the optimum incubation temperature of 40 degrees C. It is stable and retai ns its full activity after heating at 40-50 degrees C for 30 min. The activity is lost completely at 80 degrees C.