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http://dx.doi.org/10.4014/jmb.1310.10121

Cloning and Characterization of a Novel ${\alpha}$-Amylase from a Fecal Microbial Metagenome  

Xu, Bo (School of Life Science, Yunnan Normal University)
Yang, Fuya (School of Life Science, Yunnan Normal University)
Xiong, Caiyun (School of Life Science, Yunnan Normal University)
Li, Junjun (School of Life Science, Yunnan Normal University)
Tang, Xianghua (School of Life Science, Yunnan Normal University)
Zhou, Junpei (School of Life Science, Yunnan Normal University)
Xie, Zhenrong (School of Life Science, Yunnan Normal University)
Ding, Junmei (School of Life Science, Yunnan Normal University)
Yang, Yunjuan (School of Life Science, Yunnan Normal University)
Huang, Zunxi (School of Life Science, Yunnan Normal University)
Publication Information
Journal of Microbiology and Biotechnology / v.24, no.4, 2014 , pp. 447-452 More about this Journal
Abstract
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 ${\alpha}$-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 ${\alpha}$-amylase from a gastrointestinal metagenomic library.
Keywords
Amylase; metagenome; function-based screening; gastrointestinal microorganisms; cloning; characterization;
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