Optimization and purification of lipase through submerged fermentation by Alcanivorax sp. GI-CMST1 from the gut of marine fish Sardinella longicepsis

摘要

The present study was aimed at optimization and purification of lipase from Alcanivorax sp. GI-CMST. Various nutritional and physical parameters induce more production of lipase production as 2g lactose (w/v), 1.25g yeast extract (w/v), 0.5μl olive oil (v/v), pH 8.0, Temperature 50ºC and incubation time 72h. The lipase was purified by 75% (NH4)2SO4 fractionation followed by simultaneous desalting and concentrating by ultrafiltration, then chromatography as DEAE-Cellulose and Sephadex G-75 gel filtration. The molecular weight and activity of the enzyme were 42kDa as determined by crude ammonium sulphate, DEAE-Cellulose, and Sephadex G-75 gel filtration through SDS and native polyacrylamide gel electrophoresis. Detailed results on optimization and purification of lipase are discussed here with.