Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Expression, Purification, and Characterization of a Cold-adapted Lipase from Janthinobacterium sp.

Janthinobacterium sp. 유래 저온활성 lipase의 발현, 정제 및 효소 특성 연구

  • 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)
  • 박성호 (전남대학교 생물공학과, 바이오에너지 및 바이오소재 협동과정) ;
  • 박성주 (전남대학교 생물공학과, 바이오에너지 및 바이오소재 협동과정) ;
  • 최종일 (전남대학교 생물공학과, 바이오에너지 및 바이오소재 협동과정)
  • Received : 2018.02.08
  • Accepted : 2018.02.13
  • Published : 2018.03.28
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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.

본 연구에서는 극지에서 유래한 Janthinobacterium sp. PAMC25641로부터 분리한 리파아제 유전자를 클로닝 하고 과발현시켜 정제하였으며, 이 분리한 재조합 리파아제 효소의 생화학적 특성에 대해 분석하였다. 이 효소는 $15^{\circ}C$ 이하의 온도에서 장시간 활성을 유지하는 효소로서 산업적으로 활용될 가능성이 높을 것으로 기대된다.

Keywords

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