Cloning, expression and characterization of a novel esterase from a South China Sea sediment metagenome -- Journal of Oceanology and Limnology,2015,33(4):819-827

	Cite this paper:
	ZHANG Hao, LI Fuchao, CHEN Huaxin, ZHAO Jin, YAN Jinfei, JIANG Peng, LI Ronggui, ZHU Baoli. Cloning, expression and characterization of a novel esterase from a South China Sea sediment metagenome[J]. Journal of Oceanology and Limnology, 2015, 33(4): 819-827

Cloning, expression and characterization of a novel esterase from a South China Sea sediment metagenome

ZHANG Hao^1,2, LI Fuchao², CHEN Huaxin², ZHAO Jin², YAN Jinfei³, JIANG Peng², LI Ronggui¹, ZHU Baoli⁴

1 Laboratory of Microbiology, Shandong Province, Qingdao University, Qingdao 266071, China;
2 Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China

Abstract:

Lipolytic enzymes, including esterases and lipases, represent a group of hydrolases that catalyze the cleavage and formation of ester bonds. A novel esterase gene, scsEst01, was cloned from a South China Sea sediment metagenome. The scsEst01 gene consisted of 921 bp encoding 307 amino acid residues. The predicted amino acid sequence shared less than 90% identity with other lipolytic enzymes in the NCBI nonredundant protein database. ScsEst01 was successfully co-expressed in Escherichia coli BL21 (DE3) with chaperones (dnaK-dnaJ-grpE) to prevent the formation of inclusion bodies. The recombinant protein was purified on an immobilized metal ion affinity column containing chelating Sepharose charged with Ni²⁺. The enzyme was characterized using p-nitrophenol butyrate as a substrate. ScsEst01 had the highest lipolytic activity at 35℃ and pH 8.0, indicative of a meso-thermophilic alkaline esterase. ScsEst01 was thermostable at 20℃. The lipolytic activity of scsEst01 was strongly increased by Fe²⁺, Mn²⁺ and 1% Tween 80 or Tween 20.

Key words: marine sediment|metagenome|lipolytic|expression|esterase characterization

Received: 2014-06-12 Revised: 2014-08-13

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Articles by ZHANG Hao

Articles by LI Fuchao

Articles by CHEN Huaxin

Articles by ZHAO Jin

Articles by YAN Jinfei

Articles by JIANG Peng

Articles by LI Ronggui

Articles by ZHU Baoli

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