caa a22 4500 606212876 CHVBK 20210128101044.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s10295-015-1639-0 doi (NATIONALLICENCE)springer-10.1007/s10295-015-1639-0 Improvement and characterization of a hyperthermophilic glucose isomerase from Thermoanaerobacter ethanolicus and its application in production of high fructose corn syrup [Elektronische Daten] [Zhi-Qiang Liu, Wei Zheng, Jian-Feng Huang, Li-Qun Jin, Dong-Xu Jia, Hai-Yan Zhou, Jian-Miao Xu, Cheng-Jun Liao, Xin-Ping Cheng, Bao-Xing Mao, Yu-Guo Zheng] High fructose corn syrup (HFCS) is an alternative of liquid sweetener to sucrose that is isomerized by commercial glucose isomerase (GI). One-step production of 55% HFCS by thermostable GI has been drawn more and more attentions. In this study, a new hyperthermophilic GI from Thermoanaerobacter ethanolicus CCSD1 (TEGI) was identified by genome mining, and then a 1317bp fragment encoding the TEGI was synthesized and expressed in Escherichia coli BL21(DE3). To improve the activity of TEGI, two amino acid residues, Trp139 and Val186, around the active site and substrate-binding pocket based on the structural analysis and molecular docking were selected for site-directed mutagenesis. The specific activity of mutant TEGI-W139F/V186T was 2.3-fold and the value of k cat/K m was 1.86-fold as compared to the wild type TEGI, respectively. Thermostability of mutant TEGI-W139F/V186T at 90°C for 24h showed 1.21-fold extension than that of wild type TEGI. During the isomerization of glucose to fructose, the yield of fructose could maintain above 55.4% by mutant TEGI-W139F/V186T as compared to 53.8% by wild type TEGI at 90°C. This study paved foundation for the production of 55% HFCS using the thermostable TEGI. Society for Industrial Microbiology and Biotechnology, 2015 Glucose isomerase nationallicence Thermostability nationallicence High fructose corn syrup nationallicence Isomerization nationallicence Site-directed mutagenesis nationallicence Liu Zhi-Qiang Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut Zheng Wei Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut Huang Jian-Feng Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut Jin Li-Qun Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut Jia Dong-Xu Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut Zhou Hai-Yan Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut Xu Jian-Miao Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut Liao Cheng-Jun Zhejiang Huakang Pharmaceutical Co., LTD., 18 Huagong Road, Huabu Town, 324302, Kaihua, China aut Cheng Xin-Ping Zhejiang Huakang Pharmaceutical Co., LTD., 18 Huagong Road, Huabu Town, 324302, Kaihua, China aut Mao Bao-Xing Zhejiang Huakang Pharmaceutical Co., LTD., 18 Huagong Road, Huabu Town, 324302, Kaihua, China aut Zheng Yu-Guo Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/8(2015-08-01), 1091-1103 1367-5435 42:8<1091 2015 42 10295 https://doi.org/10.1007/s10295-015-1639-0 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-015-1639-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Liu Zhi-Qiang Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut NATIONALLICENCE 700 1- Zheng Wei Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut NATIONALLICENCE 700 1- Huang Jian-Feng Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut NATIONALLICENCE 700 1- Jin Li-Qun Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut NATIONALLICENCE 700 1- Jia Dong-Xu Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut NATIONALLICENCE 700 1- Zhou Hai-Yan Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut NATIONALLICENCE 700 1- Xu Jian-Miao Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut NATIONALLICENCE 700 1- Liao Cheng-Jun Zhejiang Huakang Pharmaceutical Co., LTD., 18 Huagong Road, Huabu Town, 324302, Kaihua, China aut NATIONALLICENCE 700 1- Cheng Xin-Ping Zhejiang Huakang Pharmaceutical Co., LTD., 18 Huagong Road, Huabu Town, 324302, Kaihua, China aut NATIONALLICENCE 700 1- Mao Bao-Xing Zhejiang Huakang Pharmaceutical Co., LTD., 18 Huagong Road, Huabu Town, 324302, Kaihua, China aut NATIONALLICENCE 700 1- Zheng Yu-Guo Institute of Bioengineering, Zhejiang University of Technology, 310014, Hangzhou, China aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/8(2015-08-01), 1091-1103 1367-5435 42:8<1091 2015 42 10295