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http://dx.doi.org/10.4014/jmb.1406.06071

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)
Publication Information
Journal of Microbiology and Biotechnology / v.24, no.11, 2014 , pp. 1484-1489 More about this Journal
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.
Keywords
Lipolytic enzyme; family VIII esterase; cold-adapted esterase; metagenomic library; biogas digester;
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