Cytochrome P450-catalyzed O-dealkylation coupled with photochemical NADPH regeneration

Authors

  • Sahng Ha Lee,

    1. Department of Materials Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Korea; telephone: +82-42-350-3340; fax: +82-42-350-331
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  • Yong-Chan Kwon,

    1. Department of Fine Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon 305-764, Korea; telephone: +82-42-821-5899; fax: +82-42-822-6637
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  • Dong-Myung Kim,

    Corresponding author
    1. Department of Fine Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon 305-764, Korea; telephone: +82-42-821-5899; fax: +82-42-822-6637
    • Department of Fine Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon 305-764, Korea; telephone: +82-42-821-5899; fax: +82-42-822-6637
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  • Chan Beum Park

    Corresponding author
    1. Department of Materials Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Korea; telephone: +82-42-350-3340; fax: +82-42-350-331
    • Department of Materials Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 305-701, Korea; telephone: +82-42-350-3340; fax: +82-42-350-331
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  • Sahng Ha Lee and Yong-Chan Kwon contributed equally to this work.

Abstract

Cytochrome P450 monooxygenases are multifunctional enzymes with potential applications in chemoenzymatic synthesis of complex chemicals as well as in studies of metabolism and xenobiotics. Widespread application of cytochrome P450s, however, is encumbered by the critical need for redox equivalents in their catalytic function. To overcome this limitation, we studied visible light-driven regeneration of NADPH for P450-catalyzed O-dealkylation reaction; we used eosin Y as a photosensitizing dye, triethanolamine as an electron donor, and [Cp*Rh(bpy)H2O] as an electron mediator. We analyzed catalytic activity of cell-free synthesized P450 BM3 monooxygenase variant (Y51F/F87A, BM3m2) in the presence of key components for NADPH photoregeneration. The P450-catalyzed O-dealkylation reaction sustainably maintained its turnover with the continuous supply of photoregenerated NADPH. Visible light-driven, non-enzymatic NADPH regeneration provides a new route for efficient, sustainable utilization of P450 monooxygenases. Biotechnol. Bioeng. 2013; 110: 383–390. © 2012 Wiley Periodicals, Inc.

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