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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