Studies on isolation production and purification of lipase from aeromonas Caviae Au04

Abstract

Lipases Carboxyl ester hydrolases EC3113 are enzymes which newlinehydrolyse long chain fatty acid esters in aqueous solutions Lipases also newlinecatalyse the reverse reaction from glycerol and fatty acids to form glycerides newlineby esterification in organic solvents Thus lipases are active in both aqueous newlineand nonaqueous solvent systems Most lipases are also able to catalyze newlinetransesterification and enantioselective hydrolysis reactions One of the newlinebottlenecks in the industrial applications of enzymes from extremophiles is newlinetheir low production yields Production of lipases is mainly depends on media newlinecomponents and physical conditions such as temperature pH and agitation In newlinegeneral there is no defined media available for production of lipases and so newlinedesigning appropriate production media for each microbe is required to newlinemaximize lipase production newlineA bacterium producing extracellular thermostable lipase was isolated newlinefrom milk industry waste and was identified as Aeromonas caviae AU04 newlineThe optimum conditions for production of lipase were determined by 3 step newlinestrategy Onefactoratatime PlackettBurman and Response surface newlinemethodology Suitable carbon and nitrogen source for lipase production was newlinefound to be glucose and peptone respectively For PlackettBurman design newlinenine variables were selected to study their effect on lipase production Four newlinevariables which had more effect on lipase production Glucose peptone NaCl newlineand inoculum size were selected for response surface analysis to determine newlineoptimum levels The aeration and agitation conditions were optimized in lab newlinescale Bioreactor to improve lipase production After optimization of aeration newlineand agitation conditions the maximum lipase activity of 1382 Uml was newlinefound after 7 hours of fermentation The enzyme was purified 1315fold with newline3393 recovery by ammonium sulphate precipitation and hydrophobic newlineinteraction chromatography The enzyme showed a strong tendency to newlineaggregate in solution The purified enzyme did not penetrate into stacking gel newlineand also eluted in the void volume of the column in gel pe