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

被引：6

作者：

Liu, Xinyu ^{[1
]}

Jia, Wei ^{[2
]}

An, Yi ^{[1
]}

Cheng, Kun ^{[1
]}

Wang, Mingdao ^{[1
]}

Yang, Sen ^{[1
]}

Chen, Hongge ^{[1
]}

机构：

[1] Henan Agr Univ, Minist Agr, Coll Life Sci, Key Lab Enzyme Engn Agr Microbiol, Zhengzhou 450002, Peoples R China

[2] Shanghai Acad Agr Sci, Inst Edible Fungi, Shanghai 201403, Peoples R China

来源：

JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY | 2015年 / 25卷 / 06期

关键词：

alpha-Amylase; Bacillus amyloliquefaciens; raw starch; starch-binding domain; BACILLUS-AMYLOLIQUEFACIENS; MOLECULAR-CLONING; STARCH; PURIFICATION; EXPRESSION;

D O I：

10.4014/jmb.1409.09094

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Based on its alpha-amylase activity at pH 5.0 and optimal pH of the crude enzyme, a strain (named B-5) with acid alpha-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 alpha-amylase gene of GenBank Accession No. GU318401 was cloned and expressed in Escherichia coli. The purified recombinant alpha-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 vamylase AMY-Ba in this work was different from the well-investigated J01542-type alpha-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.

引用

页码：828 / 836

页数：9

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