One-Pot Biocatalytic Double Oxidation of α-Isophorone for the Synthesis of Ketoisophorone

Michele Tavanti, Fabio Parmeggiani, J. Rubén Gómez Castellanos, Andrea Mattevi, Nicholas J. Turner

    Research output: Contribution to journalArticlepeer-review

    Abstract

    The chemical synthesis of ketoisophorone, a valuable building block of vitamins and pharmaceuticals, suffers from several drawbacks in terms of reaction conditions and selectivity. Herein, the first biocatalytic one-pot double oxidation of the readily available α-isophorone to ketoisophorone is described. Variants of the self-sufficient P450cam-RhFRed with improved activity have been identified to perform the first step of the designed cascade (regio- and enantioselective allylic oxidation of α-isophorone to 4-hydroxy-α-isophorone). For the second step, the screening of a broad panel of alcohol dehydrogenases (ADHs) led to the identification of Cm-ADH10 from Candida magnoliae. The crystal structure of Cm-ADH10 was solved and docking experiments confirmed the preferred position and geometry of the substrate for catalysis. The synthesis of ketoisophorone was demonstrated both as a one-pot two-step process and as a cascade process employing designer cells co-expressing the two biocatalysts, with a productivity of up to 1.4 g L−1 d−1.

    Original languageEnglish
    Pages (from-to)3338-3348
    Number of pages11
    JournalChemCatChem
    Volume9
    Issue number17
    Early online date4 May 2017
    DOIs
    Publication statusPublished - 2017

    Keywords

    • alcohol dehydrogenases
    • biocatalysis
    • C−H activation
    • enzyme cascades
    • P450 monooxygenases

    Research Beacons, Institutes and Platforms

    • Manchester Institute of Biotechnology

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