Characterization of Cytochrome P450 Enzymes and Their Applications in Synthetic Biology

Laura N. Jeffreys, Hazel M. Girvan, Kirsty J. Mclean, Andrew W. Munro

    Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

    Abstract

    The cytochrome P450 monooxygenase enzymes (P450s) catalyze a diverse array of chemical transformations, most originating from the insertion of an oxygen atom into a substrate that binds close to the P450 heme. The oxygen is delivered by a highly reactive heme iron-oxo species (compound I) and, according to the chemical nature of the substrate and its position in the active site, the P450 can catalyze a wide range of reactions including, e.g., hydroxylation, reduction, decarboxylation, sulfoxidation, N- and O-demethylation, epoxidation, deamination, CC bond formation and breakage, nitration, and dehalogenation. In this chapter, we describe the structural, biochemical, and catalytic properties of the P450s, along with spectroscopic and analytical methods used to characterize P450 enzymes and their redox partners. Important uses of P450 enzymes are highlighted, including how various P450s have been exploited for applications in synthetic biology.
    Original languageEnglish
    Title of host publicationEnzymes in Synthetic Biology
    Place of PublicationCambride MA, USA
    PublisherElsevier BV
    Chapter8
    Pages189-261
    Number of pages72
    Volume608
    Editionfirst
    ISBN (Print)978-0-12-815148-8
    DOIs
    Publication statusPublished - 10 Sept 2018

    Publication series

    NameMethods in Enzymology
    PublisherAcademic Press, Inc.
    ISSN (Print)0076-6879

    Keywords

    • Cytochrome P450
    • Protein expression
    • Protein purification
    • Enzyme catalysis
    • Enzyme mechanism
    • redox partner enzymes
    • UV-visible absorption spectroscopy
    • P450 quantification
    • P450 enzyme assays
    • GC-MS analysis
    • HPLC-MS
    • NMR spectroscopy
    • Protein crystallography
    • Biotechnology

    Research Beacons, Institutes and Platforms

    • Biotechnology

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