[논문]A Direct Approach for Finding Functional Lipolytic Enzymes from the Paenibacillus polymyxa Genome

JUNG, YEO-JIN; KIM, HYUNG-KWOUN; KIM, JIHYUN F.; PARK, SEUNG-HWAN; OH, TAE-KWANG; LEE, JUNG-KEE

A Direct Approach for Finding Functional Lipolytic Enzymes from the Paenibacillus polymyxa Genome 원문보기

Journal of microbiology and biotechnology, v.15 no.1, 2005년, pp.155 - 160

JUNG, YEO-JIN (Laboratory of Microbial Genomics, Korea Research Institute of Bioscience and Biotechnology) , KIM, HYUNG-KWOUN (Division of Biotechnology, The Catholic University of Korea) , KIM, JIHYUN F. (Laboratory of Microbial Genomics, Korea Research Institute of Bioscience and Biotechnology) , PARK, SEUNG-HWAN (Laboratory of Microbial Genomics, Korea Research Institute of Bioscience and Biotechnology) , OH, TAE-KWANG (Laboratory of Microbial Genomics, Korea Research Institute of Bioscience and Biotechnology) , LEE, JUNG-KEE (Laboratory of Microbial Genomics, Korea Research Institute of Bioscience and Biotechnology)

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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 pnitrophenyl 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 $30^{\circ}C$ and pH 7- 8, and was stable up to $40^{\circ}C$ and in the range of pH 5- 8. It most rapidly hydrolyzed pNPC$_6$ among various PNPesters. 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 equence, they are suggested as true lipase genes.

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