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.
|Number of pages||6|
|Journal||Journal of microbiology and biotechnology|
|Publication status||Published - 2005 Feb 1|
All Science Journal Classification (ASJC) codes
- Applied Microbiology and Biotechnology