Cloning and Expression of Ca-independent Acid α-amylase Gene

Home > Archive>Volume 37, Issue 10, 2010 >1427-1431

Cloning and Expression of Ca-independent Acid α-amylase Gene
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                        YANG JianYANG Jian
1. Guangxi Key Laboratory of Subtropical Bio-resource Conservation and Utilization, College of Life Science and Technology, Guangxi University, Nanning, Guangxi 530005, China; 2. National Non-grain Bio-energy Engineering Research Center, Guangxi Academy of Sciences, Nanning, Guangxi 530007, China
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WANG Qing-YanWANG Qing-Yan
1. Guangxi Key Laboratory of Subtropical Bio-resource Conservation and Utilization, College of Life Science and Technology, Guangxi University, Nanning, Guangxi 530005, China; 2. National Non-grain Bio-energy Engineering Research Center, Guangxi Academy of Sciences, Nanning, Guangxi 530007, China
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JIN HuiJIN Hui
1. Guangxi Key Laboratory of Subtropical Bio-resource Conservation and Utilization, College of Life Science and Technology, Guangxi University, Nanning, Guangxi 530005, China; 2. National Non-grain Bio-energy Engineering Research Center, Guangxi Academy of Sciences, Nanning, Guangxi 530007, China
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QIN YanQIN Yan
2. National Non-grain Bio-energy Engineering Research Center, Guangxi Academy of Sciences, Nanning, Guangxi 530007, China
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WANG Cheng-HuaWANG Cheng-Hua
2. National Non-grain Bio-energy Engineering Research Center, Guangxi Academy of Sciences, Nanning, Guangxi 530007, China; 3. College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing, Jiangsu 210009, China
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HUANG Ri-BoHUANG Ri-Bo
1. Guangxi Key Laboratory of Subtropical Bio-resource Conservation and Utilization, College of Life Science and Technology, Guangxi University, Nanning, Guangxi 530005, China; 2. National Non-grain Bio-energy Engineering Research Center, Guangxi Academy of Sciences, Nanning, Guangxi 530007, China
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Abstract:

The a-amylase gene cn7a was amplified by PCR from Bacillus sp. CN7 genome DNA. The recombinant plasmid pSE380-cn7a was constructed by inserting gene cn7a into expression vector pSE380 and then transformed into Escherichia coli JM109. The purified amylase CN7A showed an optimal activity at pH 5.5?6.0 and 65°C, the Km value is 3.784 g/L taking soluble starch as substrate, and the maximum velocity was determined as 101.2 mg/(L·min). Ca ion was not required for the thermal stability of CN7A.

Key words:Acid amylase, Ca-independent, Cloning and expression, pKa

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YANG Jian, WANG Qing-Yan, JIN Hui, QIN Yan, WANG Cheng-Hua, HUANG Ri-Bo. Cloning and Expression of Ca-independent Acid α-amylase Gene[J]. Microbiology China, 2010, 37(10): 1427-1431

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