Abstract

Lipases represent a group of extracellular enzymes and hold a prominent place among the enzymes. In the present research work Serratia marcescens isolated from soil was selected for lipase production. Effects of various parameters such as substrate, pH, temperature, incubation time and inoculum size were analysed for the enhanced lipase production. The optimum substrate was found to be olive oil with the concentration of 0.7 mL in 100 mL of production medium. Serratia marcescens produced maximum lipase after 48 h of incubation at 30 °C with the optimum pH as 7.0. The inoculum concentration of 4 % was more favourable for the production of lipase. The extracted lipase was purified by two steps: ultra-filtration and Carboxymethyl(CM)-Cellulose chromatography. The specific activity of the purified enzyme was determined to be 65.0 U/mg with 2.55 purification fold and yield of 67.5 %. Polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed the presence of a single band with a molecular weight of around 35 kDa. Based on Fourier transformation-infrared spectroscopy analysis the functional groups of lipase were determined and high performance liquid chromatography analysis with the retention time of 1.995 min reveals the presence of lipase in the purified extract.

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