|
KMID : 1236020180460010051
|
|
Microbiology and Biotechnology Letters
2018 Volume.46 No. 1 p.51 ~ p.58
|
|
|
Expression, Purification, and Characterization of a Cold-adapted Lipase from Janthinobacterium sp.
|
|
Park Sung-Ho
Park Seong-Ju
Choi Jong-Il
|
|
|
Abstract
|
|
|
|
The expression, purification, and characterization of cold-adapted lipase from the psychrophile, Janthinobacterium sp. were investigated. The gene encoding lipase from Janthinobacterium sp. PAMC 25641 was cloned into a pET28a(+) vector and heterologously expressed in Escherichia coli BL21 (DE3). The amino acid sequence deduced from the nucleotide sequence (930 bp) corresponded to a protein having 309 amino acid residues with a molecular weight of 32.7 kDa and a pI of 5.55. Recombinant E. coli harboring the Janthinobacterium lipase gene were induced by addition of isopropyl-β-D-thiogalactopyranoside. Ni2+-NTA affinity chromatography was used to purify the lipase, which had a specific activity of 107.9 U/mg protein. The effect of temperature and pH on the activity of lipase was measured using p-nitrophenyl octanoate as a substrate. The stability of the lipase at low temperatures indicated it is a cold-adapted enzyme. The lipase activity was increased by Na2+, Mg2+, and Mn2+, and decreased by Zn2+ and Co2+. Analysis of the lipase activity using various p-nitrophenyl esters showed a strong preference toward short acyl chains of the esters, indicating the ability of the cold-adapted lipase to hydrolyze short-chain esters.
|
|
|
KEYWORD
|
|
|
Lipase, Janthinobacterium sp., expression, characterization
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|