A Structure-Guided Switch in the Regioselectivity of a Tryptophan Halogenase

Sarah Shepherd, Binuraj Menon, Heidi Fisk, Anna-Winona Struck, Colin Levy, David Leys, Jason Micklefield

    Research output: Contribution to journalArticlepeer-review


    Flavin-dependent halogenase enzymes are potentially useful biocatalysts for the regioselectively halogenation of aromatic compounds. Haloaromatic compounds can be utilised in the synthesis and biosynthesis of pharmaceuticals and other valuable products. Here we report the first X-ray crystal structure of a tryptophan 6-halogenase (SttH), which enabled key residues that contribute to the regioselectivity in tryptophan halogenases to be identified. Structure-guided mutagenesis resulted in a triple mutant (L460F/P461E/P462T) that exhibited a complete switch in regioselectivity; with the substrate 3-indolepropionate 75% 5-chlorination was observed with the mutant in comparison with 90% 6-chlorination for the wild-type SttH. This is the first clear example of how regiocomplementary halogenase enzymes can be created from a single parent enzyme. The biocatalytic repertoire of SttH was also expanded to include a range of indolic and non-indolic substrates.
    Original languageEnglish
    Pages (from-to)821–824
    JournalChemBioChem: a European journal of chemical biology
    Early online date30 Mar 2016
    Publication statusPublished - 3 May 2016


    • Aryl halides
    • Biocatalyst
    • Biohalogenation
    • Structure guided mutagenesis
    • regioselective halogenation

    Research Beacons, Institutes and Platforms

    • Manchester Institute of Biotechnology


    Dive into the research topics of 'A Structure-Guided Switch in the Regioselectivity of a Tryptophan Halogenase'. Together they form a unique fingerprint.

    Cite this