메타게노믹스를 이용한 해양환경유래 지질분해효소 탐색

Abstract

Metagenomic libraries of various marine samples collected from a seamount near hydrothermal vent, Artic coast (Dasan station) and inter tidal flat of Kangwha island were constructed using a fosmid vector. The total size of the metagenomic library was estimated to about 8 Gb (more than 200,000 clones). The library was screened for the lipolytic activity by plating the clones on TBN plate, and as a result, eighteen positive clones forming a halo were obtained. The genes representing the lipolytic activities were cloned. The sequence analysis revealed that all of 18 genes contained an oxyanion hole (XG) and an active-site (GXSXG) consensus region, and showed a significant similarity to α/β hydrolase family, essentially esterase/lipases. In an attempt to express the esterase/lipase genes in E. coli, most of those enzymes were not produced in a soluble form. Several approaches to resolve the solubility issue are under progress.