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 Hao1,2, 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
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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