7JSD

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

7JSD の概要

• エントリーDOI: 10.2210/pdb7jsd/pdb
• 分子名称: Lysine hydroxylase, LYSINE, 2-OXOGLUTARIC ACID, ... (6 entities in total)
• 機能のキーワード: hydroxylase, non-heme iron, biosynthetic protein
• 由来する生物種: Streptomyces roseifaciens
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 121952.60

構造登録者

Kissman, E.N.,Neugebauer, M.E.,Chang, M.C.Y. (登録日: 2020-08-14, 公開日: 2021-08-25, 最終更新日: 2023-10-18)

主引用文献

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

実験手法

X-RAY DIFFRACTION (2.5 Å)