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KMID : 0545120140240111484
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Journal of Microbiology and Biotechnology
2014 Volume.24 No. 11 p.1484 ~ p.1489
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Molecular Cloning and Characterization of a Novel Cold-Adapted Family VIII Esterase from a Biogas Slurry Metagenomic Library
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Xiaojie Cheng
Xuming Wang
Tianlei Qiu
Mei Yuan
Jianguang Sun
Junlian Gao
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Abstract
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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°C (43% activity remained), with the optimal temperature at 20°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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KEYWORD
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Lipolytic enzyme, Family VIII esterase, Cold-adapted esterase, Metagenomic library, Biogas digester
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