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Synthetic cascades are enabled by combining biocatalysts with artificial metalloenzymes

Nature Chemistry volume 5, pages 9399 (2013) | Download Citation

Abstract

Enzymatic catalysis and homogeneous catalysis offer complementary means to address synthetic challenges, both in chemistry and in biology. Despite its attractiveness, the implementation of concurrent cascade reactions that combine an organometallic catalyst with an enzyme has proven challenging because of the mutual inactivation of both catalysts. To address this, we show that incorporation of a d6-piano stool complex within a host protein affords an artificial transfer hydrogenase (ATHase) that is fully compatible with and complementary to natural enzymes, thus enabling efficient concurrent tandem catalysis. To illustrate the generality of the approach, the ATHase was combined with various NADH-, FAD- and haem-dependent enzymes, resulting in orthogonal redox cascades. Up to three enzymes were integrated in the cascade and combined with the ATHase with a view to achieving (i) a double stereoselective amine deracemization, (ii) a horseradish peroxidase-coupled readout of the transfer hydrogenase activity towards its genetic optimization, (iii) the formation of L-pipecolic acid from L-lysine and (iv) regeneration of NADH to promote a monooxygenase-catalysed oxyfunctionalization reaction.

  • Compound C10H11N

    1-Methyl-3,4-dihydroisoquinoline

  • Compound C10H13N

    1-Methyl-1,2,3,4-tetrahydroisoquinoline

  • Compound C10H17N

    2-Cyclohexyl-1-pyrroline

  • Compound C10H19N

    2-Cyclohexylpyrrolidine

  • Compound C10H14N2O

    4-(Methylamino)-1-(pyridin-3-yl)butan-1-one

  • Compound C10H14N2

    3-(1-Methylpyrrolidin-2-yl)pyridine

  • Compound C10H16N2O

    4-(Methylamino)-1-(pyridin-3-yl)butan-1-ol

  • Compound C6H14N2O2

    L-Lysine

  • Compound C6H11NO2

    Piperidine-2-carboxylic acid

  • Compound C12H10O

    [1,1'-Biphenyl]-2-ol

  • Compound C12H10O2

    [1,1'-Biphenyl]-2,3-diol

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Acknowledgements

This work was supported by the Marie Curie Initial Training Network (Biotrains FP7-ITN-238531). T.R.W. acknowledges financial support from the SNF (Schweizerische Nationalfonds, grant no. 200020_126366) and the National Centre of Competence in Research Nanosciences. N.J.T. acknowledges the Royal Society for a Wolfson Research Merit Award. F.H. thanks A. Schmid (Dortmund University of Technology) for the kind provision of HbpA. The authors also thank M. Corbett, S. Willies and K. Malone for helpful advice and materials, R. Pfalzberger for help with the graphic material, and Umicore for a precious metal loan.

Author information

Affiliations

  1. Department of Chemistry, University of Basel, Spitalstrasse 51, CH-4056 Basel, Switzerland

    • V. Köhler
    • , Y. M. Wilson
    • , M. Dürrenberger
    • , T. Quinto
    • , L. Knörr
    • , D. Häussinger
    •  & T. R. Ward
  2. School of Chemistry, University of Manchester, Manchester Institute of Biotechnology, 131 Princess Street, Manchester M1 7DN, UK

    • D. Ghislieri
    •  & N. J. Turner
  3. Department of Biotechnology, Delft University of Technology, Julianalaan 136, 2628BL Delft, The Netherlands

    • E. Churakova
    •  & F. Hollmann

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Contributions

V.K., F.H., N.T. and T.W. conceived the catalytic cascades. V.K., F.H., N.T. and T.W. supervised the project. V.K., Y.W., M.D., D.G., E.C. and T.Q. performed the experiments. V.K., Y.W., M.D., D.G., E.C., T.Q., F.H., N.T. and T.W. analysed the data. V.K., F.H., N.T. and T.W. co-wrote the paper. V.K., Y.W., M.D., D.G. and L.K. contributed materials. D.H. analysed the conversion of 13C-labelled lysine by 2D NMR.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to F. Hollmann or N. J. Turner or T. R. Ward.

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DOI

https://doi.org/10.1038/nchem.1498

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