期刊
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
卷 24, 期 6, 页码 771-780出版社
KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
DOI: 10.4014/jmb.1311.11071
关键词
Esterase; marine metagenome; salt tolerance
资金
- National High Technology Research and Development Program of China [2012AA092105]
- Scientific Research Foundation for Returned Scholars, Ministry of Education of China
- Introduction Project of Academic and Technology Leaders in Anhui University [32030066]
- Innovative Research Team Program of 211 Project in Anhui University
A putative lipolytic enzyme gene, named as est9x, was obtained from a marine microbial metagenome of the South China Sea. Sequence analysis showed that Est9X shares lower than 27% sequence identities with the characterized lipolytic enzymes, but possesses a catalytic triad highly conserved in lipolytic enzymes of the alpha/beta hydrolase superfamily. By phylogenetic tree construction, Est9X was grouped into a new lipase/esterase family. To understand Est9X protein in depth, it was recombinantly expressed, purified, and biochemically characterized. Within potential hydrolytic activities, only lipase/esterase activity was detected for Est9X, confirming its identity as a lipolytic enzyme. When using p-nitrophenol esters with varying lengths of fatty acid as substrates, Est9X exhibited the highest activity to the C2 substrate, indicating it is an esterase. The optimal activity of Est9X occurred at a temperature of 65 degrees C, and Est9X was pretty stable below the optimum temperature. Distinguished from other salt-tolerant esterases, Est9X's activity was tolerant to and even promoted by as high as 4 M NaCl. Our results imply that Est9X is a unique esterase and could be a potential candidate for industrial application under extreme conditions.
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