Cloning, Expression and Biochemical Characterization of the Recombinant α-amylase from Bacillus subtilis YX48

(E-pub Abstract Ahead of Print)

Author(s): Yan Shan, Junjie Shang*, Dongfang Zhang, Yinshan Cui, Yi Wang, Jie Zhu, Yongkai Ma, Pengfei Song, Kunhao Qin, Xiuling Ji, Yunlin Wei*, Lijun Wu

Journal Name: Current Proteomics


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Abstract:

Background: Amylase used in the market is mostly medium-temperature enzyme or high-temperature enzyme and has poor enzyme activity under low-temperature environment. Acid α-amylase can be used to develop digestion additives in the pharmaceutical and healthcare industries. The amino acid sequence and structural differences among α-amylases obtained from various organisms are high enough to confer interesting biochemical diversity to the enzymes. However, low- temperature (0-50℃) amylase, with an optimum temperature and heat sensitivity, has a greater potential value than medium (50-80℃) and high (80-110℃) temperature amylases.

Methodology: The gene amy48 from encoding extracellular α-amylase in Bacillus subtilis YX48 was successfully cloned into the pET30a (+) vector and expressed in Escherichia coli BL21 (DE3) for biochemical characterization.

Results and Conclusion: The molecular weight of α-amylase was 75 kDa. The activity of α-amylase was not affected by Ca2+, and Amy48 had the best activity at pH 5.0 and 37℃. AMY48 has high stability over a narrow pH and temperature range (5.0-8.0 and 30-45℃). Amylase activity was strongly inhibited by Zn2+, Mn2+, Cu2+, and Fe2+ ions, but Na+, K+, and Co2+ ions stimulate its activity slightly. The purified enzyme showed gradually reduced activity in the presence of detergents. However, it was remarkably stable against EDTA and urea.

Keywords: α-amylase, Bacillus subtilis, YX48, Characterization, Organic, Solvents

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Article Details

Published on: 26 July, 2021
(E-pub Abstract Ahead of Print)
DOI: 10.2174/1570164618666210726161428
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