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http://dx.doi.org/10.4014/jmb.1409.09094

Screening, Gene Cloning, and Characterizations of an Acid-Stable α-Amylase  

Liu, Xinyu (Key Laboratory of Enzyme Engineering of Agricultural Microbiology, Ministry of Agriculture, College of Life Sciences, Henan Agricultural University)
Jia, Wei (Institute of Edible Fungi, Shanghai Academy of Agricultural Sciences)
An, Yi (Key Laboratory of Enzyme Engineering of Agricultural Microbiology, Ministry of Agriculture, College of Life Sciences, Henan Agricultural University)
Cheng, Kun (Key Laboratory of Enzyme Engineering of Agricultural Microbiology, Ministry of Agriculture, College of Life Sciences, Henan Agricultural University)
Wang, Mingdao (Key Laboratory of Enzyme Engineering of Agricultural Microbiology, Ministry of Agriculture, College of Life Sciences, Henan Agricultural University)
Yang, Sen (Key Laboratory of Enzyme Engineering of Agricultural Microbiology, Ministry of Agriculture, College of Life Sciences, Henan Agricultural University)
Chen, Hongge (Key Laboratory of Enzyme Engineering of Agricultural Microbiology, Ministry of Agriculture, College of Life Sciences, Henan Agricultural University)
Publication Information
Journal of Microbiology and Biotechnology / v.25, no.6, 2015 , pp. 828-836 More about this Journal
Abstract
Based on its α-amylase activity at pH 5.0 and optimal pH of the crude enzyme, a strain (named B-5) with acid α-amylase production was screened. The B-5 strain was identified as Bacillus amyloliquefaciens through morphological, physiological, and biochemical characteristics analysis, as well as 16S rDNA phylogenetic analysis. Its α-amylase gene of GenBank Accession No. GU318401 was cloned and expressed in Escherichia coli. The purified recombinant α-amylase AMY-Ba showed the optimal pH of 5.0, and was stable at a pH range of 4.0-6.0. When hydrolyzing soluble starch, amylose, and amylopectin, AMY-Ba released glucose and maltose as major end products. The α-amylase AMY-Ba in this work was different from the well-investigated J01542-type α-amylase which also came from B. amyloliquefaciens. AMY-Ba exhibited notable adsorption and hydrolysis ability towards various raw starches. Structure analysis of AMY-Ba suggested the presence of a new starch-binding domain at its C-terminal region.
Keywords
α-Amylase; Bacillus amyloliquefaciens; raw starch; starch-binding domain;
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