8ZOP

Structure of FasV in complex with FAD

8ZOP の概要

• エントリーDOI: 10.2210/pdb8zop/pdb
• 分子名称: Halogenase, FLAVIN-ADENINE DINUCLEOTIDE, CHLORIDE ION, ... (4 entities in total)
• 機能のキーワード: naphthacemycins, flavin-dependent halogenases, fasv, biosynthetic protein
• 由来する生物種: Streptomyces sp.
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 46716.26

構造登録者

Ma, X.Y.,Chen, Q.Q. (登録日: 2024-05-29, 公開日: 2025-04-02)

主引用文献

Hu, Y.,Peng, S.Y.,Ma, X.,Chen, H.,Nie, Q.Y.,He, J.B.,Chen, Q.,Zhou, Q.,Lu, X.H.,Hua, Q.,Yang, D.,Liang, Y.,Ma, M.,Tang, G.L.
Functional Characterization and Molecular Basis of a Multi-Site Halogenase in Naphthacemycin Biosynthesis.
Angew.Chem.Int.Ed.Engl., 64:e202418843-e202418843, 2025

PubMed Abstract: Halogenases are spurring a growing interest in the fields of biosynthesis and biocatalysis. Although various halogenases have been identified in numerous natural product biosynthetic pathways, the mechanisms for multiple halogenations and site-selectivity remain largely unclear. In this study, we biochemically characterized FasV, a flavin-dependent halogenase (FDH) that catalyzes five successive chlorinations in the biosynthesis of the naphthacene-containing aromatic polyketide naphthacemycin. This multiple halogenation reaction was elucidated to occur in an orderly fashion, as evidenced by enzyme kinetics, time-course assays, and computational simulations. Crystallographic analyses and mutagenesis studies revealed previously unrecognized amino acid residues, including T53, L81, F93, and I212, that are crucial for controlling regioselectivity and substrate specificity. Based on this, a I212T mutant was generated to exclusively catalyze selective monohalogenation. We propose a novel dual-activation mechanism and demonstrate that the larger binding pocket of FasV makes it a valuable biocatalyst for other substrates with diverse structures. Therefore, this study provides new insight into multi-site polyhalogenases and highlights the potential for engineering FasV-like FDHs for biocatalytic applications.

PubMed: 39612320
DOI: 10.1002/anie.202418843

実験手法

X-RAY DIFFRACTION (1.5 Å)