잠시만 기다려 주세요. 로딩중입니다.

Isolation and Characterization of a Novel Triolein Selective Lipase from Soil Environmental Genes

Microbiology and Biotechnology Letters 2020년 48권 4호 p.480 ~ 490
임희경, 한예진, Hahm Moon-Sun, Park Soo-Youl, 황인택,
소속 상세정보
임희경 ( Lim Hee-Kyung ) - Korea Research Institute of Chemical Technology
한예진 ( Han Ye-Jin ) - Korea Research Institute of Chemical Technology
 ( Hahm Moon-Sun ) - Bioprogen Co. Ltd.
 ( Park Soo-Youl ) - Korea Research Institute of Chemical Technology
황인택 ( Hwang In-Taek ) - Korea Research Institute of Chemical Technology

Abstract


A novel lipase gene, Lip-1420, was isolated from a metagenomic library constructed from reed marsh from Mt. Jumbong in Korea, comprising 112,500 members of recombinant plasmids. The DNA sequence of Lip- 1420-subclone (5,513 bp) was found to contain at least 11 ORFs according to the GenBank database. The ORF-3 gene was inserted into the pET21a plasmid containing the C-terminal 6-His tag and transformed into E. coli BL21(DE3) to express the recombinant lipase protein. Lip-1420 was purified using a fast protein liquid chromatography system. The gene was registered in GenBank (MH628529). The values of Km and Vmax were determined as 0.268 mM and 1.821 units, respectively, at 40℃ and pH 8.0, using p-nitrophenyl palmitate as the substrate. This lipase belongs to family IV taxonomically because it has conserved HGGG and GDSAG motifs in the constitutive amino acid sequence. According to the predicted structural model, the binding sites are represented by residues H78, G81, D150, S151, A152, V181, and D236. Finally, Lip-1420 showed triolein selectivity for methanolysis between triolein (18:1) and tristearin (18:0) substrates. Further study of the selective mechanism and structure-function relationship of this new lipase could be useful for more practical applications.

키워드

Lipase; rhizosphere; metagenomics; 3D-structure; triolein selectivity

원문 및 링크아웃 정보

등재저널 정보