PROCESS FACILITATED ENHANCEMENT OF PRODUCTION AND PURIFICATION OF LIPASE USING BACILLUS SPP

Abstract

newline quotLipase is the extensively used key enzyme for various newlinebiotechnological applications in wide spectrum of industries. The process newlineoriented feeding strategies for higher production of lipase was objective of the newlinestudy. Range of carbon sources were used for the higher production of lipase, newlineamong them germinated maize oil was chosen as the best one. Feeding newlinestrategies like quasi steady state, pH stat and DO stat were employed. newlineEventually it was established that the DO stat based fed batch fermentation newlinegave maximum in terms of both enzymatic units and productivity. The yield newlinein DO stat fed batch fermentation was seven fold higher when compared to newlinebatch fermentation. newlineThe optimum temperature and pH was arrived at from a range of newlinetemperature and pH. Further the activity of lipase was observed by using newlinedifferent pH and temperatures. It was found that at pH 6.5 and temperature at newline37oC the lipase enzyme was most active. The purification of the lipase done newlineby ammonium sulphate precipitation and two stage chromatographic methods. newlineThe crude lipase was precipitated with saturated concentration of ammonium newlinesulphate and the high salt concentration was reduced by dialysis. With the newlinehelp of size exclusion chromatography and SDS-PAGE, molecular weight of newlinethe protein was identified which was nearly 25 kDa.quot newline