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A growth selection system for the directed evolution of amine-forming or converting enzymes

Author

Listed:
  • Shuke Wu

    (University of Greifswald
    Huazhong Agricultural University)

  • Chao Xiang

    (University of Greifswald)

  • Yi Zhou

    (Huazhong Agricultural University)

  • Mohammad Saiful Hasan Khan

    (University of Greifswald)

  • Weidong Liu

    (University of Greifswald
    Chinese Academy of Sciences)

  • Christian G. Feiler

    (Helmholtz-Zentrum Berlin für Materialien und Energie)

  • Ren Wei

    (University of Greifswald)

  • Gert Weber

    (Helmholtz-Zentrum Berlin für Materialien und Energie)

  • Matthias Höhne

    (University of Greifswald)

  • Uwe T. Bornscheuer

    (University of Greifswald)

Abstract

Fast screening of enzyme variants is crucial for tailoring biocatalysts for the asymmetric synthesis of non-natural chiral chemicals, such as amines. However, most existing screening methods either are limited by the throughput or require specialized equipment. Herein, we report a simple, high-throughput, low-equipment dependent, and generally applicable growth selection system for engineering amine-forming or converting enzymes and apply it to improve biocatalysts belonging to three different enzyme classes. This results in (i) an amine transaminase variant with 110-fold increased specific activity for the asymmetric synthesis of the chiral amine intermediate of Linagliptin; (ii) a 270-fold improved monoamine oxidase to prepare the chiral amine intermediate of Cinacalcet by deracemization; and (iii) an ammonia lyase variant with a 26-fold increased activity in the asymmetric synthesis of a non-natural amino acid. Our growth selection system is adaptable to different enzyme classes, varying levels of enzyme activities, and thus a flexible tool for various stages of an engineering campaign.

Suggested Citation

  • Shuke Wu & Chao Xiang & Yi Zhou & Mohammad Saiful Hasan Khan & Weidong Liu & Christian G. Feiler & Ren Wei & Gert Weber & Matthias Höhne & Uwe T. Bornscheuer, 2022. "A growth selection system for the directed evolution of amine-forming or converting enzymes," Nature Communications, Nature, vol. 13(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-35228-y
    DOI: 10.1038/s41467-022-35228-y
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    References listed on IDEAS

    as
    1. Lara Sellés Vidal & James W. Murray & John T. Heap, 2021. "Versatile selective evolutionary pressure using synthetic defect in universal metabolism," Nature Communications, Nature, vol. 12(1), pages 1-15, December.
    2. Kevin M. Esvelt & Jacob C. Carlson & David R. Liu, 2011. "A system for the continuous directed evolution of biomolecules," Nature, Nature, vol. 472(7344), pages 499-503, April.
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    Cited by:

    1. Chenghai Sun & Gen Lu & Baoming Chen & Guangjun Li & Ya Wu & Yannik Brack & Dong Yi & Yu-Fei Ao & Shuke Wu & Ren Wei & Yuhui Sun & Guifa Zhai & Uwe T. Bornscheuer, 2024. "Direct asymmetric synthesis of β-branched aromatic α-amino acids using engineered phenylalanine ammonia lyases," Nature Communications, Nature, vol. 15(1), pages 1-13, December.
    2. Enrico Orsi & Lennart Schada von Borzyskowski & Stephan Noack & Pablo I. Nikel & Steffen N. Lindner, 2024. "Automated in vivo enzyme engineering accelerates biocatalyst optimization," Nature Communications, Nature, vol. 15(1), pages 1-14, December.

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