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Co-immobilized Alcohol Dehydrogenase and Glucose Dehydrogenase with Resin Extraction for Continuous Production of Chiral Diaryl Alcohol

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Abstract

Ni2+-functionalized porous ceramic/agarose composite beads (Ni-NTA Cerose) can be used as carrier materials to immobilize enzymes harboring a metal affinity tag. Here, a 6×His-tag fusion alcohol dehydrogenase Mu-S5 and glucose dehydrogenase from Bacillus megaterium (BmGDH) were co-immobilized on Ni-NTA Cerose to construct a packed bed reactor (PBR) for the continuous synthesis of the chiral intermediate (S)-(4-chlorophenyl)-(pyridin-2-yl) methanol ((S)-CPMA) NADPH recycling, and in situ product adsorption was achieved simultaneously by assembling a D101 macroporous resin column after the PBR. Using an optimum enzyme activity ratio of 2:1 (Mu-S5: BmGDH) and hydroxypropyl-β-cyclodextrin as co-solvent, a space-time yield of 1560 g/(L·d) could be achieved in the first three days at a flow rate of 5 mL/min and substrate concentration of 10 mM. With simplified selective adsorption and extraction procedures, (S)-CPMA was obtained in 84% isolated yield.

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Abbreviations

ADH:

Alcohol dehydrogenase

GDH:

Glucose dehydrogenase

FDH:

Formate dehydrogenase

CPMK:

(4-Chlorophenyl)(pyridine-2-yl)ketone

(S)-CPMA:

(S)-(4-Chlorophenyl)-(pyridin-2-yl) methanol

DNPH:

2,4-Dinitrophenylhydrazine

PBR:

Packed bed reactor

Cerose:

Ceramic/agarose composite beads

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Funding

This work was supported by the National Key R&D Program [2018YFA0901700], the National Natural Science Foundation of China [21907040, 21776112, 22077054], China Postdoctoral Science Foundation [2019 M651703], the National First-Class Discipline Program of Light Industry Technology and Engineering [LITE2018-07], and the Program of Introducing Talents of Discipline to Universities [111-2-06].

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Authors and Affiliations

  1. Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, Jiangsu, China

    Jieyu Zhou, Yanfei Wu, Qingye Zhang, Guochao Xu & Ye Ni

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  1. Jieyu Zhou
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  2. Yanfei Wu
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  3. Qingye Zhang
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  4. Guochao Xu
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Contributions

Jieyu Zhou: Conceptualization, investigation, methodology, writing-original draft. Yanfei Wu: Data curation. Qingye Zhang: Data curation, formal analysis. Guochao Xu: Writing—review and editing. Ye Ni: Supervision, writing—review and editing, project administration, funding acquisition.

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Correspondence to Ye Ni.

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Zhou, J., Wu, Y., Zhang, Q. et al. Co-immobilized Alcohol Dehydrogenase and Glucose Dehydrogenase with Resin Extraction for Continuous Production of Chiral Diaryl Alcohol. Appl Biochem Biotechnol 193, 2742–2758 (2021). https://doi.org/10.1007/s12010-021-03561-5

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