7JSD

Hydroxylase homolog of BesD with Fe(II), alpha-ketoglutarate, and lysine

Summary for 7JSD

• Entry DOI: 10.2210/pdb7jsd/pdb
• Descriptor: Lysine hydroxylase, LYSINE, 2-OXOGLUTARIC ACID, ... (6 entities in total)
• Functional Keywords: hydroxylase, non-heme iron, biosynthetic protein
• Biological source: Streptomyces roseifaciens
• Total number of polymer chains: 4
• Total formula weight: 121952.60

Authors

Kissman, E.N.,Neugebauer, M.E.,Chang, M.C.Y. (deposition date: 2020-08-14, release date: 2021-08-25, Last modification date: 2023-10-18)

Primary citation

Neugebauer, M.E.,Kissman, E.N.,Marchand, J.A.,Pelton, J.G.,Sambold, N.A.,Millar, D.C.,Chang, M.C.Y.
Reaction pathway engineering converts a radical hydroxylase into a halogenase.
Nat.Chem.Biol., 18:171-179, 2022

PubMed Abstract: Fe/α-ketoglutarate (Fe/αKG)-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/αKG-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.

PubMed: 34937913
DOI: 10.1038/s41589-021-00944-x

Experimental method

X-RAY DIFFRACTION (2.5 Å)