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Cloning and Characterization of a Novel α-Amylase from a Fecal Microbial MetagenomeS
Cloning and Characterization of a Novel α-Amylase from a Fecal Microbial MetagenomeS
Journal of Microbiology and Biotechnology. 2014. Apr, 24(4): 447-452
Copyright © 2014, The Korean Society For Microbiology And Biotechnology
  • Received : November 05, 2013
  • Accepted : January 02, 2014
  • Published : April 30, 2014
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About the Authors
Bo Xu
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming 650500, China
Fuya Yang
School of Life Science, Yunnan Normal University, Kunming 650500, China
Caiyun Xiong
School of Life Science, Yunnan Normal University, Kunming 650500, China
Junjun Li
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming 650500, China
Xianghua Tang
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming 650500, China
Junpei Zhou
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming 650500, China
Zhenrong Xie
School of Life Science, Yunnan Normal University, Kunming 650500, China
Junmei Ding
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming 650500, China
Yunjuan Yang
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming 650500, China
Zunxi Huang
Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming 650500, China
huangzunxi@163.com

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 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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Acknowledgements
This work was supported by the National Natural Science Foundation of China (30960165, 31360268, and 31160229).
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