Enhanced catalytic efficiency and thermostability of glucose isomerase from Thermoanaerobacter ethanolicus via site-directed mutagenesis

被引:9
|
作者
Jin, Li-Qun [1 ,2 ,3 ]
Jin, Yi-Ting [1 ,2 ,3 ]
Zhang, Jing-Wei [1 ,2 ,3 ]
Liu, Zhi-Qiang [1 ,2 ,3 ]
Zheng, Yu-Guo [1 ,2 ,3 ]
机构
[1] Zhejiang Univ Technol, Natl & Local Joint Engn Res Ctr Biomfg Chiral Che, Hangzhou 310014, Zhejiang, Peoples R China
[2] Zhejiang Univ Technol, Minist Educ, Engn Res Ctr Bioconvers & Biopurificat, Hangzhou 310014, Zhejiang, Peoples R China
[3] Zhejiang Univ Technol, Coll Biotechnol & Bioengn, Key Lab Bioorgan Synth Zhejiang Prov, Hangzhou 310014, Zhejiang, Peoples R China
基金
国家重点研发计划;
关键词
Glucose isomerase; Thermostability; Site-specific mutagenesis; High fructose corn syrup; BIOCHEMICAL-CHARACTERIZATION; HIGH-LEVEL; EXPRESSION; IMPROVEMENT;
D O I
10.1016/j.enzmictec.2021.109931
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
Glucose isomerase (GI) is a key enzyme in the preparation of high fructose corn syrup (HFCS). In this study, a mutant TEGI-M-L38 M/V137 L (TEGI-M2) of glucose isomerase (TEGI-M) originated from Thermoanaerobacter ethanalicus CCSD1 was obtained by site-directed mutagenesis. The TEGI-M2 showed an optimal activity at 85 degrees C and pH 6.5 with the divalent cations Co2+ and Mg2+. The structural differences between TEGI-M and TEGI-M2 were investigated based on the homology modeling and molecular docking, to elucidate the mechanism of improvement in the enzymatic properties. Compared with the original enzyme, the TEGI-M2 showed a 2.0-fold increased enzyme activity and a decreased K-m from 234.2 mM to 85.9 mM. Finally, the application of mutant TEGI-M2 in HFCS one-step biosynthesis was attempted, resulting in a D-fructose yield of 67.3 %, which was 14.3 % higher than that of TEGI-M. This improved catalytic performance of TEGI-M2 was of great importance for the industrial preparation of D-fructose in one-step process.
引用
收藏
页数:9
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