期刊
ENZYME AND MICROBIAL TECHNOLOGY
卷 152, 期 -, 页码 -出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.enzmictec.2021.109931
关键词
Glucose isomerase; Thermostability; Site-specific mutagenesis; High fructose corn syrup
资金
- National Key R&D Program of China [2020YFA0908400]
- Fundamental Research Funds for the Provincial Universities of Zhejiang [RF-C2019005]
Glucose isomerase mutant TEGI-M2 showed optimal activity at high temperature and neutral pH, with structural differences compared to the original enzyme investigated through molecular docking. TEGI-M2 exhibited increased enzyme activity and decreased Km, resulting in higher D-fructose yield in one-step biosynthesis of HFCS. This improved catalytic performance of TEGI-M2 is significant for industrial production of D-fructose.
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.
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