Structure and biocatalytic scope of thermophilic flavin-dependent halogenase and flavin reductase enzymes

Binuraj R. K. Menon, Jonathan Latham, Mark S. Dunstan, Eileen Brandenburger, Ulrike Klemstein, David Leys, Chinnan Karthikeyan, Michael F. Greaney, Sarah A. Shepherd, Jason Micklefield*

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

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    Abstract

    Flavin-dependent halogenase (Fl-Hal) enzymes have been shown to halogenate a range of synthetic as well as natural aromatic compounds. The exquisite regioselectively of Fl-Hal enzymes can provide halogenated building blocks which are inaccessible using standard halogenation chemistries. Consequently, Fl-Hal are potentially useful biocatalysts for the chemoenzymatic synthesis of pharmaceuticals and other valuable products, which are derived from haloaromatic precursors. However, the application of Fl-Hal enzymes, in vitro, has been hampered by their poor catalytic activity and lack of stability. To overcome these issues, we identified a thermophilic tryptophan halogenase (Th-Hal), which has significantly improved catalytic activity and stability, compared with other Fl-Hal characterised to date. When used in combination with a thermostable flavin reductase, Th-Hal can efficiently halogenate a number of aromatic substrates. X-ray crystal structures of Th-Hal, and the reductase partner (Th-Fre), provide insights into the factors that contribute to enzyme stability, which could guide the discovery and engineering of more robust and productive halogenase biocatalysts.

    Original languageEnglish
    Pages (from-to)9354-9361
    Number of pages8
    JournalOrganic and Biomolecular Chemistry
    Volume14
    Issue number39
    Early online date6 Sept 2016
    DOIs
    Publication statusPublished - 21 Oct 2016

    Keywords

    • UKRI
    • BBSRC
    • BB/K00199X/1

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