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KMID : 0545120140240040447
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Journal of Microbiology and Biotechnology
2014 Volume.24 No. 4 p.447 ~ p.452
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Cloning and Characterization of a Novel α-Amylase from a Fecal Microbial Metagenome
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Bo Xu
Fuya Yang
Caiyun Xiong
Junjun Li
Xianghua Tang
Junpei Zhou
Zhenrong Xie
Junmei Ding
Yunjuan Yang
Zunxi Huang
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Abstract
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To isolate novel and useful microbial enzymes from uncultured gastrointestinal microorganisms, a fecal microbial metagenomic library of the pygmy loris was constructed. The library was screened for amylolytic activity, and 8 of 50,000 recombinant clones showed amylolytic activity. Subcloning and sequence analysis of a positive clone led to the identification a novel gene (amyPL) coding for α-amylase. AmyPL was expressed in Escherichia coli BL21 (DE3) and the purified AmyPL was enzymatically characterized. This study is the first to report the molecular and biochemical characterization of a novel α-amylase from a gastrointestinal metagenomic library.
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KEYWORD
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amylase, metagenome, function-based screening, gastrointestinal microorganisms, cloning, characterization
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