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Brazilian Journal of Microbiology

Print version ISSN 1517-8382

Abstract

BORKAR, Prita S.; BODADE, Ragini G.; RAO, Srinivasa R.  and  KHOBRAGADE, C.N.. Purification and characterization of extracellular lipase from a new strain: Pseudomonas aeruginosa SRT 9. Braz. J. Microbiol. [online]. 2009, vol.40, n.2, pp. 358-366. ISSN 1517-8382.  http://dx.doi.org/10.1590/S1517-83822009000200028.

An extra cellular lipase was isolated and purified from the culture broth of Pseudomonas aeruginosa SRT 9 to apparent homogeneity using ammonium sulfate precipitation followed by chromatographic techniques on phenyl Sepharose CL- 4B and Mono Q HR 5/5 column, resulting in a purification factor of 98 fold with specific activity of 12307.8 U/mg. The molecular weight of the purified lipase was estimated by SDS-PAGE to be 29 kDa with isoelectric point of 4.5. Maximum lipase activity was observed in a wide range of temperature and pH values with optimum temperature of 55ºC and pH 6.9. The lipase preferably acted on triacylglycerols of long chain (C14-C16) fatty acids. The lipase was inhibited strongly by EDTA suggesting the enzyme might be metalloprotein. SDS and metal ions such as Hg2+, Zn2+, Cu2+, Ag2+ and Fe2+ decreased the lipase activity remarkedly. Its marked stability and activity in organic solvents suggest that this lipase is highly suitable as a biotechnological tool with a variety of applications including organo synthetic reactions and preparation of enantiomerically pure pharmaceuticals. The Km and Vmax value of the purified enzyme for triolein hydrolysis were calculated to be 1.11 mmol/L and 0.05 mmol/L/minrespectively.

Keywords : Pseudomonas aeruginosa SRT9; extra cellular lipases; purification; Michaelis constant.

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