A direct approach for finding functional lipolytic enzymes from the Paenibacillus polymyxa genome

Yeo Jin Jung, Hyung Kwoun Kim, Jihyun F. Kim, Seung Hwan Park, Tae Kwang Oh, Jung Kee Lee

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

A direct approach was used to retrieve active lipases from Paenibacillus polymyxa genome databases. Twelve putative lipase genes were tested using a typical lipase sequence rule built on the basis of a consensus sequence of a catalytic triad and oxyanion hole. Among them, six genes satisfied the sequence rule and had similarity (about 25%) with known bacterial lipases. To obtain the six lipase proteins, lipase genes were expressed in E. coli cells and lipolytic activities were measured by using tributyrin plate and p-nitrophenyl caproate. One of them, contig 160-26, was expressed as a soluble and active form in E. coli cell. After purifying on Ni-NTA column, its detailed biochemical properties were characterized. It had a maximum hydrolytic activity at 30°C and pH 7-8, and was stable up to 40°C and in the range of pH 5-8. It most rapidly hydrolyzed pNPC6 among various PNP-esters. The other contigs were expressed more or less as soluble forms, although no lipolytic activities were detected. As they have many conserved regions with lipase 160-26 as well as other bacterial lipases throughout their sequence, they are suggested as true lipase genes.

Original languageEnglish
Pages (from-to)155-160
Number of pages6
JournalJournal of microbiology and biotechnology
Volume15
Issue number1
Publication statusPublished - 2005 Feb 1

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All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology

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