Get access

Photoenzymatic Reduction of C[DOUBLE BOND]C Double Bonds

Authors

  • Maria Mifsud Grau,

    1. Delft University of Technology, Department of Biotechnology, Biocatalysis and Organic Chemistry Group, Julianalaan 136, 2628 BL Delft, The Netherlands, Fax: (+31)-152-781-415; phone: (+31)-152-781-957
    Search for more papers by this author
  • John C. van der Toorn,

    1. Delft University of Technology, Department of Biotechnology, Biocatalysis and Organic Chemistry Group, Julianalaan 136, 2628 BL Delft, The Netherlands, Fax: (+31)-152-781-415; phone: (+31)-152-781-957
    Search for more papers by this author
  • Linda G. Otten,

    1. Delft University of Technology, Department of Biotechnology, Biocatalysis and Organic Chemistry Group, Julianalaan 136, 2628 BL Delft, The Netherlands, Fax: (+31)-152-781-415; phone: (+31)-152-781-957
    Search for more papers by this author
  • Peter Macheroux,

    1. Graz University of Technology, Institute of Biochemistry, Petersgasse 12, 8010 Graz, Austria
    Search for more papers by this author
  • Andreas Taglieber,

    1. Firmenich SA, Corporate R&D, Biotechnology & Bioengineering, Route des Jeunes 1, 1211 Geneva 8, Switzerland
    Search for more papers by this author
  • Felipe E. Zilly,

    1. Max-Planck-Institut für Kohlenforschung, Kaiser-Wilhelm-Platz 1, 45470 Mülheim an der Ruhr, Germany
    Search for more papers by this author
  • Isabel W. C. E. Arends,

    1. Delft University of Technology, Department of Biotechnology, Biocatalysis and Organic Chemistry Group, Julianalaan 136, 2628 BL Delft, The Netherlands, Fax: (+31)-152-781-415; phone: (+31)-152-781-957
    Search for more papers by this author
  • Frank Hollmann

    1. Delft University of Technology, Department of Biotechnology, Biocatalysis and Organic Chemistry Group, Julianalaan 136, 2628 BL Delft, The Netherlands, Fax: (+31)-152-781-415; phone: (+31)-152-781-957
    Search for more papers by this author

Abstract

A simplified procedure for cell-free biocatalytic reductions of conjugated C[DOUBLE BOND]C double bonds using old yellow enzymes (OYEs) is reported. Instead of indirectly regenerating YqjM (an OYE homologue from B. subtilis) or NemA (N-ethylmaleimide reductase from E. coli) via regeneration of reduced nicotinamide cofactors, we demonstrate that direct regeneration of catalytically active reduced flavins is an efficient and convenient approach. Reducing equivalents are provided from simple sacrificial electron donors such as ethylenediaminetetraacetate (EDTA), formate, or phosphite via photocatalytic oxidation. This novel photoenzymatic reaction scheme was characterized. Up to 65% rates of the NADH-driven reaction were obtained while preserving enantioselectivity. The chemoselectivity of the novel approach was exclusive. Even when using crude cell extracts as biocatalyst preparations, only C[DOUBLE BOND]C bond reduction was observed while ketone and aldehyde groups remained unaltered. Overall, a simple and practical approach for photobiocatalytic reductions is presented.

Ancillary