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Structure-Based Insight into the Asymmetric Bioreduction of the C[DOUBLE BOND]C Double Bond of α,β-Unsaturated Nitroalkenes by Pentaerythritol Tetranitrate Reductase

Authors

  • Helen S. Toogood,

    1. Manchester Interdisciplinary Biocentre, Faculty of Life Sciences, University of Manchester, 131 Princess Street, Manchester M1 7DN, U.K. Fax: (+44)-161-306-8918; phone: (+44)-161-306-5152
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    • The first two authors contributed equally to this work.

  • Anna Fryszkowska,

    1. Manchester Interdisciplinary Biocentre, Department of Chemistry, University of Manchester, 131 Princess Street, Manchester M1 7DN, U.K.
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    • The first two authors contributed equally to this work.

  • Victoria Hare,

    1. Manchester Interdisciplinary Biocentre, Faculty of Life Sciences, University of Manchester, 131 Princess Street, Manchester M1 7DN, U.K. Fax: (+44)-161-306-8918; phone: (+44)-161-306-5152
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  • Karl Fisher,

    1. Manchester Interdisciplinary Biocentre, School of Chemical Engineering and Analytical Sciences, University of Manchester, 131 Princess Street, Manchester M1 7DN, U.K.
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  • Anna Roujeinikova,

    1. Manchester Interdisciplinary Biocentre, Faculty of Life Sciences, University of Manchester, 131 Princess Street, Manchester M1 7DN, U.K. Fax: (+44)-161-306-8918; phone: (+44)-161-306-5152
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  • David Leys,

    1. Manchester Interdisciplinary Biocentre, Faculty of Life Sciences, University of Manchester, 131 Princess Street, Manchester M1 7DN, U.K. Fax: (+44)-161-306-8918; phone: (+44)-161-306-5152
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  • John M. Gardiner,

    1. Manchester Interdisciplinary Biocentre, Department of Chemistry, University of Manchester, 131 Princess Street, Manchester M1 7DN, U.K.
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  • Gill M. Stephens,

    1. Manchester Interdisciplinary Biocentre, School of Chemical Engineering and Analytical Sciences, University of Manchester, 131 Princess Street, Manchester M1 7DN, U.K.
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  • Nigel S. Scrutton

    1. Manchester Interdisciplinary Biocentre, Faculty of Life Sciences, University of Manchester, 131 Princess Street, Manchester M1 7DN, U.K. Fax: (+44)-161-306-8918; phone: (+44)-161-306-5152
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Abstract

Biocatalytic reduction of α- or β-alkyl-β-arylnitroalkenes provides a convenient and efficient method to prepare chiral substituted nitroalkanes. Pentaerythritol tetranitrate reductase (PETN reductase) from Enterobacter cloacae st. PB2 catalyses the reduction of nitroolefins such as 1-nitrocyclohexene (1) with steady state and rapid reaction kinetics comparable to other old yellow enzyme homologues. Furthermore, it reduces 2-aryl-1-nitropropenes (4a–d) to their equivalent (S)-nitropropanes 9a–d. The enzyme shows a preference for the (Z)-isomer of substrates 4a–d, providing almost pure enantiomeric products 9a–d (ees up to>99%) in quantitative yield, whereas the respective (E)-isomers are reduced with lower enantioselectivity (63–89% ee) and lower product yields. 1-Aryl-2-nitropropenes (5a, b) are also reduced efficiently, but the products (R)-10 have lower optical purities. The structure of the enzyme complex with 1-nitrocyclohexene (1) was determined by X-ray crystallography, revealing two substrate-binding modes, with only one compatible with hydride transfer. Models of nitropropenes 4 and 5 in the active site of PETN reductase predicted that the enantioselectivity of the reaction was dependent on the orientation of binding of the (E)- and (Z)-substrates. This work provides a structural basis for understanding the mechanism of asymmetric bioreduction of nitroalkenes by PETN reductase.

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