Isolation, Identification and Characterization of a Lipase Producing Pseudomonas

Abstract

Lipases are hydrolytic enzymes and catalyze the hydrolysis of long-chain triacylglycerols into glycerol and fatty acid. Lipases are produced by plants, animals and microorganisms including bacteria and fungi. However microbial lipases, especially from bacteria, are more useful than their plant and animal derivatives because of several important properties. The primary goals of this research work is to isolate and identify a lipase producing Pseudomonas species from waste water samples collected from Dir lower, Peshawar and Kohat Khyber pakhtunkhwa Pakistan. The isolated bacteria were identified as Pseudomonas biochemically. The other purposes of this study are production, partial purification, characterization of lipase activity at different pH and incubation time, production and determination of molecular weight analysis. The lipase was partially purified up to 30% saturation using ammonium sulphate precipitation. Purity of lipases was cheeked by SDS-PAGE, showing the same banding pattern of all the lipases and the molecular weight were determined approximately 50kDA by comparing with protein marker bands. Spectrophotometric lipase assay was used for enzyme characterization. All the 7 isolates shows maximum activity at pH 7 after 48 hours of incubation and 37°C. Among all the isolates, isolate HSWPC shows highest activity of 110.11U /ml at pH 7 after 48 hours of incubation and 37°C. Pseudomonas lipases are widely used in food industry, detergent, paper, textile, leather and pharmaceutical industries because of their stability, selectivity and broad substrate specificity.