Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Molecular Cloning and Characterization of a Novel Cold-Adapted Family VIII Esterase from a Biogas Slurry Metagenomic Library

  • Cheng, Xiaojie (Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Science/Beijing Municipal Key Laboratory of Agricultural Gene Resources and Biotechnology) ;
  • Wang, Xuming (Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Science/Beijing Municipal Key Laboratory of Agricultural Gene Resources and Biotechnology) ;
  • Qiu, Tianlei (Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Science/Beijing Municipal Key Laboratory of Agricultural Gene Resources and Biotechnology) ;
  • Yuan, Mei (Key Laboratory of Microbial Resources, Ministry of Agriculture/Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences) ;
  • Sun, Jianguang (Key Laboratory of Microbial Resources, Ministry of Agriculture/Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences) ;
  • Gao, Junlian (Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Science/Beijing Municipal Key Laboratory of Agricultural Gene Resources and Biotechnology)
  • Received : 2014.06.30
  • Accepted : 2014.07.22
  • Published : 2014.11.28
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Abstract

A novel esterase gene, est01, was successfully unearthed from a biogas digester microbiota metagenomic library. The 1,194 bp est01 gene encodes a protein of 44,804 Da (designated Est01). The amino acid sequence of Est01 shows only moderate (33%) identity to a lipase/esterase. Phylogenetic analysis and biochemical characterization confirmed that Est01 is a new member of family VIII esterases. The purified Est01 from recombinant Escherichia coli BL21 (DE3) showed high hydrolytic activity against short-chain fatty acid esters, suggesting that it is a typical carboxylesterase rather than a lipase. Furthermore, the Est01 was even active at $10^{\circ}C$ (43% activity remained), with the optimal temperature at $20^{\circ}C$, and had a broad pH range from 5.0 to 10.0, with the optimal pH of 8.0. These properties suggest that Est01 is a cold-adaptive esterase and could have good potential for low-temperature hydrolysis application.

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