Structural highlights
Publication Abstract from PubMedFe(II)/alpha-ketoglutarate (Fe(II)/alphaKG)-dependent enzymes offer a promising biocatalytic platform for halogenation chemistry owing to their ability to functionalize unactivated C-H bonds. However, relatively few radical halogenases have been identified to date, limiting their synthetic utility. Here, we report a strategy to expand the palette of enzymatic halogenation by engineering a reaction pathway rather than substrate selectivity. This approach could allow us to tap the broader class of Fe(II)/alphaKG-dependent hydroxylases as catalysts by their conversion to halogenases. Toward this goal, we discovered active halogenases from a DNA shuffle library generated from a halogenase-hydroxylase pair using a high-throughput in vivo fluorescent screen coupled to an alkyne-producing biosynthetic pathway. Insights from sequencing halogenation-active variants along with the crystal structure of the hydroxylase enabled engineering of a hydroxylase to perform halogenation with comparable activity and higher selectivity than the wild-type halogenase, showcasing the potential of harnessing hydroxylases for biocatalytic halogenation. Reaction pathway engineering converts a radical hydroxylase into a halogenase.,Neugebauer ME, Kissman EN, Marchand JA, Pelton JG, Sambold NA, Millar DC, Chang MCY Nat Chem Biol. 2022 Feb;18(2):171-179. doi: 10.1038/s41589-021-00944-x. Epub 2021, Dec 22. PMID:34937913[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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