Cloning, expression, and characterization of a lipolytic enzyme gene (lip8) from Pseudomonas aeruginosa LST-03

Journal of Molecular Microbiology and Biotechnology
H OginoHaruo Ishikawa

Abstract

A lipolytic enzyme gene (lip8) was cloned from organic solvent-tolerant Pseudomonas aeruginosa LST-03 and sequenced. In the sequenced nucleotides, an open reading frame consisting of 1,173 nucleotides and encoding 391 amino acids was found. Lip8 is considered to belong to the family VIII of lipolytic enzymes whose serine in the consensus sequence of -Ser-Xaa-Xaa-Lys- acts as catalytic nucleophile. The gene was expressed in Escherichia coli and purified by a combination of ammonium sulfate fractionation and hydrophobic interaction and ion-exchange chromatographies to homogeneity on SDS-PAGE analysis. The optimum temperature and heat stability of Lip8 were not as high as those of Lip3 and LST-03 lipase, two other lipolytic enzymes from the same strain. Addition of glycerol to a solution containing Lip8 stabilized this enzyme. By measuring the activities against various triacylglycerols and fatty acid methyl esters having carbon chains of different lengths, Lip8 was categorized as an esterase which has higher activities against fatty acid methyl esters with short-chain fatty acids.

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Citations

Jul 25, 2009·Applied Microbiology and Biotechnology·Bhawna Madan, Prashant Mishra
Jul 28, 2007·Extremophiles : Life Under Extreme Conditions·Hiroyasu OginoHaruo Ishikawa
Aug 19, 2007·Journal of Molecular Microbiology and Biotechnology·Konanani J RashamuseDon A Cowan
Jun 14, 2012·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Bum Han RyuT Doohun Kim
Jan 26, 2010·Biochemical and Biophysical Research Communications·Yong Ho KimHoon Kim

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