Improvement and characterization of a hyperthermophilic glucose isomerase from Thermoanaerobacterethanolicus and its application in production of high fructose corn syrup View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2015-06-16

AUTHORS

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

ABSTRACT

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 1317 bp 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 kcat/Km was 1.86-fold as compared to the wild type TEGI, respectively. Thermostability of mutant TEGI-W139F/V186T at 90 °C for 24 h 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. More... »

PAGES

1091-1103

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s10295-015-1639-0

DOI

http://dx.doi.org/10.1007/s10295-015-1639-0

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1004959534

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/26077737


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36 schema:description 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 1317 bp 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 kcat/Km was 1.86-fold as compared to the wild type TEGI, respectively. Thermostability of mutant TEGI-W139F/V186T at 90 °C for 24 h 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.
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44 Km
45 Trp139
46 Val186
47 acid residues
48 active site
49 activity
50 alternative
51 amino acid residues
52 analysis
53 applications
54 attention
55 bp fragment
56 characterization
57 coli
58 corn syrup
59 docking
60 extension
61 foundation
62 fragments
63 fructose
64 fructose corn syrup
65 genome mining
66 glucose
67 glucose isomerase
68 high fructose corn syrup
69 improvement
70 isomerase
71 isomerization
72 isomerization of glucose
73 kcat/Km
74 liquid sweetener
75 mining
76 molecular docking
77 more attention
78 mutagenesis
79 one-step production
80 pocket
81 production
82 residues
83 site-directed mutagenesis
84 sites
85 specific activity
86 structural analysis
87 study
88 substrate-binding pocket
89 sweeteners
90 syrup
91 thermostability
92 thermostable glucose isomerase
93 values
94 yield
95 yield of fructose
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