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Mining and Characterization of Thermophilic Glucose Isomerase Based on Virtual Probe Technology

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Abstract

Fructose, which is produced by the isomerization of glucose isomerase, is a crucial precursor for the biosynthesis of rare sugars. In this study, thermophilic glucose isomerases (GI) from Caldicellulosiruptor acetigenus (CAGI), Thermoanaerobacter thermocopriae (TTGI), and Thermotoga petrophila (TPGI) were screened from GenBank database by a virtual probe and were successfully expressed in Escherichia coli BL21(DE3). The results of characterization demonstrated that the optimal pH for CAGI and TTGI were 8.0 and were maintained at 80% in a slightly acidic environment. The relative residual activities of CAGI and TTGI were found to be 40.6% and 52.6%, respectively, following an incubation period of 24 h at 90 ℃. Furthermore, CAGI and TTGI exhibited superior catalytic performance that their reaction equilibrium both reached only after an hour at 85 ℃ with 200 g/L glucose, and the highest conversion rates were 54.2% and 54.1%, respectively. This study identifies competitive enzyme candidates for fructose production in the industry with appreciable cost reduction.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This work was supported by the Innovation Demonstration Special Project of Henan Province in China (grant number 201111110100) and Major Science and Technology Projects for Public Welfare of Henan Province in China (grant number 201300110300).

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Authors and Affiliations

  1. School of Biological Engineering, Henan University of Technology, Zhengzhou, 450001, People’s Republic of China

    Yu-Qi Dong, Long Pan & Ji-Hong Huang

  2. The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou, 310014, People’s Republic of China

    Yu-Qi Dong, Ji-Dong Shen, Zhi-Qiang Liu & Yu-Guo Zheng

  3. Henan Provincial Key Laboratory of Biological Processing and Nutritional Function of Wheat, Zhengzhou, 450001, People’s Republic of China

    Long Pan & Ji-Hong Huang

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Contributions

Yu-Qi Dong: methodology, software, validation, investigation, and writing—original draft. Ji-Dong Shen: methodology, software, validation, investigation, and writing—original draft. Long Pan: validation and investigation. Ji-Hong Huang: visualization and supervision. Zhi-Qiang Liu: visualization and supervision. Yu-Guo Zheng: supervision.

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Correspondence to Ji-Hong Huang or Zhi-Qiang Liu.

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Dong, YQ., Shen, JD., Pan, L. et al. Mining and Characterization of Thermophilic Glucose Isomerase Based on Virtual Probe Technology. Appl Biochem Biotechnol 195, 4399–4413 (2023). https://doi.org/10.1007/s12010-023-04349-5

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