Development of a thioredoxin based cofactor regeneration system for NADPH‐dependent oxidoreductases

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  • Aarhus University
  • Technische Universität Dresden
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Original languageEnglish
Article numbere202101625
JournalCHEMCATCHEM
Volume14
Issue number7
Early online date18 Jan 2022
Publication statusPublished - 7 Apr 2022

Abstract

Nicotinamide cofactor-dependent oxidoreductases have become a valuable tool for the synthesis of high value chiral compounds. The feasibility of biocatalytic processes involving these enzymes stands and falls with the efficiency of the regeneration of cofactors. In this study, we describe a novel NADPH regeneration method based on the natural thioredoxin electron delivery system. Thioredoxin 1 (Trx1) and thioredoxin reductase (TR) from Thermus thermophilus were characterized for the dithiol-dependent reduction of NADP +, revealing good catalytic activities and a particularly remarkable thermostability. The TR/Trx1 system was then coupled with two representative NADPH-dependent oxidoreductases, alcohol dehydrogenase and cyclohexanone monooxygenase. Reaction conditions for both systems were optimized for reaction yield and selectivity. The results demonstrate the feasibility of the TR/Trx1-system for its application as NADPH regeneration system.

Keywords

    Biocatalysis, Cofactor regeneration, Enzymatic cascades, NADPH, Reductase (TR)

ASJC Scopus subject areas

Cite this

Development of a thioredoxin based cofactor regeneration system for NADPH‐dependent oxidoreductases. / Zhang, Ningning; Müller, Beatrice; Ørtoft Kirkeby, Tanja et al.
In: CHEMCATCHEM, Vol. 14, No. 7, e202101625, 07.04.2022.

Research output: Contribution to journalArticleResearchpeer review

Zhang N, Müller B, Ørtoft Kirkeby T, Kara S, Loderer C. Development of a thioredoxin based cofactor regeneration system for NADPH‐dependent oxidoreductases. CHEMCATCHEM. 2022 Apr 7;14(7):e202101625. Epub 2022 Jan 18. doi: 10.1002/cctc.202101625
Zhang, Ningning ; Müller, Beatrice ; Ørtoft Kirkeby, Tanja et al. / Development of a thioredoxin based cofactor regeneration system for NADPH‐dependent oxidoreductases. In: CHEMCATCHEM. 2022 ; Vol. 14, No. 7.
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abstract = "Nicotinamide cofactor-dependent oxidoreductases have become a valuable tool for the synthesis of high value chiral compounds. The feasibility of biocatalytic processes involving these enzymes stands and falls with the efficiency of the regeneration of cofactors. In this study, we describe a novel NADPH regeneration method based on the natural thioredoxin electron delivery system. Thioredoxin 1 (Trx1) and thioredoxin reductase (TR) from Thermus thermophilus were characterized for the dithiol-dependent reduction of NADP +, revealing good catalytic activities and a particularly remarkable thermostability. The TR/Trx1 system was then coupled with two representative NADPH-dependent oxidoreductases, alcohol dehydrogenase and cyclohexanone monooxygenase. Reaction conditions for both systems were optimized for reaction yield and selectivity. The results demonstrate the feasibility of the TR/Trx1-system for its application as NADPH regeneration system. ",
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note = "Funding Information: S.K. and N.Z. thank to Deutsche Forschungsgemeinschaft (DFG), grant agreement No KA 4399/3‐1. S.K. thanks to Aarhus University Research Foundation (AUFF) for the financial support. Open Access funding enabled and organized by Projekt DEAL.",
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AU - Zhang, Ningning

AU - Müller, Beatrice

AU - Ørtoft Kirkeby, Tanja

AU - Kara, Selin

AU - Loderer, Christoph

N1 - Funding Information: S.K. and N.Z. thank to Deutsche Forschungsgemeinschaft (DFG), grant agreement No KA 4399/3‐1. S.K. thanks to Aarhus University Research Foundation (AUFF) for the financial support. Open Access funding enabled and organized by Projekt DEAL.

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N2 - Nicotinamide cofactor-dependent oxidoreductases have become a valuable tool for the synthesis of high value chiral compounds. The feasibility of biocatalytic processes involving these enzymes stands and falls with the efficiency of the regeneration of cofactors. In this study, we describe a novel NADPH regeneration method based on the natural thioredoxin electron delivery system. Thioredoxin 1 (Trx1) and thioredoxin reductase (TR) from Thermus thermophilus were characterized for the dithiol-dependent reduction of NADP +, revealing good catalytic activities and a particularly remarkable thermostability. The TR/Trx1 system was then coupled with two representative NADPH-dependent oxidoreductases, alcohol dehydrogenase and cyclohexanone monooxygenase. Reaction conditions for both systems were optimized for reaction yield and selectivity. The results demonstrate the feasibility of the TR/Trx1-system for its application as NADPH regeneration system.

AB - Nicotinamide cofactor-dependent oxidoreductases have become a valuable tool for the synthesis of high value chiral compounds. The feasibility of biocatalytic processes involving these enzymes stands and falls with the efficiency of the regeneration of cofactors. In this study, we describe a novel NADPH regeneration method based on the natural thioredoxin electron delivery system. Thioredoxin 1 (Trx1) and thioredoxin reductase (TR) from Thermus thermophilus were characterized for the dithiol-dependent reduction of NADP +, revealing good catalytic activities and a particularly remarkable thermostability. The TR/Trx1 system was then coupled with two representative NADPH-dependent oxidoreductases, alcohol dehydrogenase and cyclohexanone monooxygenase. Reaction conditions for both systems were optimized for reaction yield and selectivity. The results demonstrate the feasibility of the TR/Trx1-system for its application as NADPH regeneration system.

KW - Biocatalysis

KW - Cofactor regeneration

KW - Enzymatic cascades

KW - NADPH

KW - Reductase (TR)

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