Catalytic Activity Improvement of Acid-stable Alpha-Amylase from Hyperthermophilic Thermococcus siculi HJ21 by Site-directed Mutagenesis

doi:10.7506/spkx1002-6630-201115034

FOOD SCIENCE ›› 2011, Vol. 32 ›› Issue (15): 148-152.doi: 10.7506/spkx1002-6630-201115034

• Bioengineering • Previous Articles Next Articles

Catalytic Activity Improvement of Acid-stable Alpha-Amylase from Hyperthermophilic Thermococcus siculi HJ21 by Site-directed Mutagenesis

YAO Ting1,2，LI Hua-zhong1，FANG Yao-wei2，LU Zhao-xin3，WANG Shu-jun2,*，JIAO Yu-liang2，LIU Shu2

(1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China ； 2. School of Marine Science and Technology, Huaihai Institute of Technology, Lianyungang 222005, China； 3. College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China)

Online:2011-08-15 Published:2011-07-26

Abstract

Abstract: Based on the analysis of hyperthermophilic acid-stable α-amylase gene from Thermococcus siculi HJ21 (TSA), site-directed mutagenesis of six amino acid residues could improve the catalytic activity of α-amylase. Five mutants with higher catalytic activity were identified as K98R, A109V, Y121S, V125L and Y184H, respectively. Subsequent recombination of the mutants produced a mutant enzyme named as TSAM-23. Compared with TSA, the catalytic activity of TSAM-23 was enhanced significantly. The catalytic activity and thermal stability of TSAM-23 at 90 ℃were increased by 3.75 times and 2.4%, respectively. TSAM-23 had an optimal pH of 5.0, which revealed a decrease of 0.5 when compared with TSA.

Key words: α-amylase, catalytic activity, site-directed mutagenesis

CLC Number:

YAO Ting，LI Hua-zhong，FANG Yao-wei，LU Zhao-xin，WANG Shu-jun，JIAO Yu-liang，LIU Shu. Catalytic Activity Improvement of Acid-stable Alpha-Amylase from Hyperthermophilic Thermococcus siculi HJ21 by Site-directed Mutagenesis[J]. FOOD SCIENCE, 2011, 32(15): 148-152.

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Catalytic Activity Improvement of Acid-stable Alpha-Amylase from Hyperthermophilic Thermococcus siculi HJ21 by Site-directed Mutagenesis

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