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http://dx.doi.org/10.4014/mbl.1802.02003

Expression, Purification, and Characterization of a Cold-adapted Lipase from Janthinobacterium sp.  

Park, Sung-ho (Department of Biotechnology and Bioengineering, Interdisciplinary Program for Bioenergy & Biomaterials, Chonnam National University)
Park, Seong-ju (Department of Biotechnology and Bioengineering, Interdisciplinary Program for Bioenergy & Biomaterials, Chonnam National University)
Choi, Jong-il (Department of Biotechnology and Bioengineering, Interdisciplinary Program for Bioenergy & Biomaterials, Chonnam National University)
Publication Information
Microbiology and Biotechnology Letters / v.46, no.1, 2018 , pp. 51-58 More about this Journal
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-${\beta}$-D-thiogalactopyranoside. $Ni^{2+}$-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 $Na^{2+}$, $Mg^{2+}$, and $Mn^{2+}$, and decreased by $Zn^{2+}$ and $Co^{2+}$. 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.
Keywords
Lipase; Janthinobacterium sp.; expression; characterization;
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