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Characterization and Application of a Novel Glucose Dehydrogenase with Excellent Organic Solvent Tolerance for Cofactor Regeneration in Carbonyl Reduction

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Abstract

An efficient cofactor regeneration system has been developed to provide a hydride source for the preparation of optically pure alcohols by carbonyl reductase-catalyzed asymmetric reduction. This system employed a novel glucose dehydrogenase (BcGDH90) from Bacillus cereus HBL-AI. The gene encoding BcGDH90 was found through the genome-wide functional annotation. Homology-built model study revealed that BcGDH90 was a homo-tetramer, and each subunit was composed of βD-αE-αF-αG-βG motif, which was responsible for substrate binding and tetramer formation. The gene of BcGDH90 was cloned and expressed in Escherichia coli. The recombinant BcGDH90 exhibited maximum activity of 45.3 U/mg at pH 9.0 and 40 °C. BcGDH90 showed high stability in a wide pH range of 4.0–10.0 and was stable after the incubation at 55 °C for 5 h. BcGDH90 was not a metal ion-dependent enzyme, but Zn2+ could seriously inhibit its activity. BcGDH90 displayed excellent tolerance to 90% of acetone, methanol, ethanol, n-propanol, and isopropanol. Furthermore, BcGDH90 was applied to regenerate NADPH for the asymmetric biosynthesis of (S)-(+)-1-phenyl-1,2-ethanediol ((S)-PED) from hydroxyacetophenone (2-HAP) with high concentration, which increased the final efficiency by 59.4%. These results suggest that BcGDH90 is potentially useful for coenzyme regeneration in the biological reduction.

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Funding

This research was funded by the Natural Science Foundation (grant number 3210120332), the Central Government Guided Local Science and Technology Development Fund of Hebei Province (grant number 216Z2603G), and the Bureau of Science and Technology of Hebei Province (CN) (grant number 20322901D).

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Authors and Affiliations

  1. College of Chemistry and Materials Science, Key Laboratory of Chemical Biology of Hebei Province, Laboratory of Medicinal Chemistry and Molecular Diagnosis of the Ministry of Education, Institute of Life Science and Green Development, Hebei University, Baoding, 071002, China

    Xiaozheng Li, Xinyue Li, Dexu Liu, Mengnan Han, Wei Li & Honglei Zhang

  2. College of Life Science, Microbial Technology Innovation Center for Feed of Hebei Province, Hebei Agricultural University, Baoding, 071001, China

    Junpo Jiang

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  1. Xiaozheng Li
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Contributions

All authors contributed to the study conception and design. Conceptualization, investigation, experimentation, and original draft preparation were performed by Xiaozheng Li. Conceptualization, manuscript reviewing, and supervision were performed by Honglei Zhang. Conceptualization and editing the manuscript were performed by Wei Li. Material preparation, data collection, and analysis were performed by Junpo Jiang, Xinyue Li, Dexu Liu, and Mengnan Han. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Li, X., Jiang, J., Li, X. et al. Characterization and Application of a Novel Glucose Dehydrogenase with Excellent Organic Solvent Tolerance for Cofactor Regeneration in Carbonyl Reduction. Appl Biochem Biotechnol 195, 7553–7567 (2023). https://doi.org/10.1007/s12010-023-04432-x

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