Molecular cloning and characterization of a novel cold-adapted family VIII esterase from a biogas slurry metagenomic library.
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.
Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Science / Beijing Municipal Key Laboratory of Agricultural Gene Resources and Biotechnology, Beijing 100097, P.R. China.
Xiaojie
X
Cheng
Xuming
X
Wang
Tianlei
T
Qiu
Mei
M
Yuan
Jianguang
J
Sun
Junlian
J
Gao
D000595:Amino Acid Sequence; D056804:Biofuels; D003001:Cloning, Molecular; D003080:Cold Temperature; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D004950:Esterases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015723:Gene Library; D006863:Hydrogen-Ion Concentration; D054892:Metagenome / Q000235:genetics*; D008969:Molecular Sequence Data; D010802:Phylogeny; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D013696:Temperature
D056804:Biofuels; D011994:Recombinant Proteins; D004950:Esterases