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KMID : 0613820120220030306
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Journal of Life Science
2012 Volume.22 No. 3 p.306 ~ p.311
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Cloning, Expression, and Purification of a Lipase from Psychrotrophic Pseudomonas mandelii
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Kim Jun-Sung
Lee Chang-Woo
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Abstract
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A gene encoding a lipase, lipT, was cloned from the psychrotrophic bacterium Pseudomonas mandelii and sequenced. An open reading frame of 1,686 bp was found that encodes a polypeptide consisting of 562 amino acid residues. Sequence analysis revealed a Gly-His-Ser-Leu-Gly sequence, which matches the consensus Gly-X-Ser-X-Gly motif conserved among lipolytic enzymes. The recombinant LipT protein was predominantly expressed as inclusion bodies in Escherichia coli and subsequently purified by nickel-chelate affinity chromatography. A small fraction of LipT was refolded, and the subsequent LipT exhibited substrate preferences for p-nitrophenyl butyrate (C4) and p-nitrophenyl octanoate (C8).
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KEYWORD
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Lipase, Pseudomonas fluorescens, Pseudomonas mandelii, Psychrotrophic Bacterium
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