8W72

Crystal structure of a P450 enzyme DmlH that catalyze intramolecular phenol coupling in the biosynthesis of cihanmycins

8W72 の概要

• エントリーDOI: 10.2210/pdb8w72/pdb
• 関連するPDBエントリー: 8W6Z
• 分子名称: Cytochrome P450, PROTOPORPHYRIN IX CONTAINING FE (2 entities in total)
• 機能のキーワード: p450 enzyme, intramolecular phenol coupling, cihanmycins, biosynthetic protein
• 由来する生物種: Streptomyces sp. DM14
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 47475.04

構造登録者

Fang, C.,Zhang, L.,Zhu, Y.,Zhang, C. (登録日: 2023-08-30, 公開日: 2024-06-12, 最終更新日: 2024-12-25)

主引用文献

Fang, C.,Zhang, L.,Wang, Y.,Xiong, W.,Yan, Z.,Zhang, W.,Zhang, Q.,Wang, B.,Zhu, Y.,Zhang, C.
Discovery and Biosynthesis of Cihanmycins Reveal Cytochrome P450-Catalyzed Intramolecular C-O Phenol Coupling Reactions.
J.Am.Chem.Soc., 2024

PubMed Abstract: Cinnamoyl-containing nonribosomal peptides (CCNPs) constitute a unique family of natural products. The enzyme mechanism for the biaryl phenol coupling reaction of the bicyclic CCNPs remains unclear. Herein, we report the discovery of two new arabinofuranosylated bicyclic CCNPs cihanmycins (CHMs) A () and B () from DSM 45679 and the identification of the CHM biosynthetic gene cluster ( BGC) by heterologous expression in SBT18 to afford CHMs C () and D (). The structure of was confirmed by X-ray diffraction analysis. Three cytochrome P450 enzyme (CYP)-encoding genes , , and were individually inactivated in the heterologous host to produce CHMs E (), F (), and G (), respectively. The structures of and indicated that Cih26 was responsible for the hydroxylation and epoxidation of the cinnamoyl moiety, and Cih32 should catalyze the β-hydroxylation of three amino acid residues. Cih33 and its homologues DmlH and EpcH were biochemically verified to convert CHM G () with a monocyclic structure to a bicyclic skeleton of CHM C () through an intramolecular C-O phenol coupling reaction. The substrate -bound crystal structure of DmlH not only established the structure of , which was difficult for NMR analysis for displaying anomalous splitting signals, but also provided the binding mode of macrocyclic peptides recognized by these intramolecular C-O coupling CYPs. In addition, computational studies revealed a water-mediated diradical mechanism for the C-O phenol coupling reaction. These findings have shed important mechanistic insights into the CYP-catalyzed phenol coupling reactions.

PubMed: 38842938
DOI: 10.1021/jacs.4c02841

実験手法

X-RAY DIFFRACTION (3.25 Å)