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Bulletin of Fermentation Science and Technology >

2015 , Vol. 44 >Issue 4: 19 - 23

DOI: https://doi.org/10.16774/j.cnki.issn.1674-2214.2015.04.007

Genome mining, expression and characterization of a hyperthermophilic glucose isomerase from Thermotoga petrophila RKU-1

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  • 1. Zhejiang Huakang Pharmaceutical Co., Ltd., Quzhou 324302, China;
    2. Institute of Bioengineering, Zhejiang University of Technology, Hangzhou 310014, China
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Abstract

A hyperthermophilic glucose isomerase from Thermotoga petrophila RKU-1 (TPGI) was screened by genome mining from NCBI database. The gene of TPGI was synthesized and expressed as a soluble protein in E. coli BL21 (DE3). The recombinant enzyme was purified by Ni-NTA affinity chromatography. The SDS-PAGE analysis of the purified TPGI revealed a single band with a molecular weight of around 50.8 kDa. The specific activity of the recombinant TPGI to D -
glucose was 19.8 U/mg. The enzyme exhibited its maximum activity at 85 ℃, pH 7.0 and showed significant dependence on Co2+. The Km and Vmax to substrate D-glucose were 373 mmol/L and 13.4 U/ mg, respectively. The yield of D-fructose reached 53.8% and the isomerization gradually maintained
equilibrium after 4 h under the optimum conditions.

Key words： genome mining; glucose isomerase; expression; enzymatic properties

Cite this article

LIAO Chengjun, ZHENG Wei, HUANG Jianfeng, LIU Chenglong, LIU Zhiqiang, CHENG Xinping, MAO Baojun, CHEN Deshui . Genome mining, expression and characterization of a hyperthermophilic glucose isomerase from Thermotoga petrophila RKU-1[J]. Bulletin of Fermentation Science and Technology, 2015 , 44(4) : 19 -23 . DOI: 10.16774/j.cnki.issn.1674-2214.2015.04.007

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