6EM0

Crystal Structure of 2-hydroxybiphenyl 3-monooxygenase M321A from Pseudomonas azelaica

6EM0 の概要

• エントリーDOI: 10.2210/pdb6em0/pdb
• 分子名称: 2-hydroxybiphenyl-3-monooxygenase, FLAVIN-ADENINE DINUCLEOTIDE (3 entities in total)
• 機能のキーワード: 2-hydroxybiphenyl 3-monooxygenase (hbpa), flavin monooxygenases2-hydroxybiphenyl, site-specific mutagenesis, oxidoreductase
• 由来する生物種: Pseudomonas nitroreducens
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 128142.47

構造登録者

Deri, B.,Bregman-Cohen, A.,Pazy Benhar, Y.,Fishman, A. (登録日: 2017-10-01, 公開日: 2018-01-10, 最終更新日: 2024-01-17)

主引用文献

Bregman-Cohen, A.,Deri, B.,Maimon, S.,Pazy, Y.,Fishman, A.
Altering 2-Hydroxybiphenyl 3-Monooxygenase Regioselectivity by Protein Engineering for the Production of a New Antioxidant.
Chembiochem, 19:583-590, 2018

PubMed Abstract: 2-Hydroxybiphenyl 3-monooxygenase is a flavin-containing NADH-dependent aromatic hydroxylase that oxidizes a broad range of 2-substituted phenols. In order to modulate its activity and selectivity, several residues in the active site pocket were investigated by saturation mutagenesis. Variant M321A demonstrated altered regioselectivity by oxidizing 3-hydroxybiphenyl for the first time, thus enabling the production of a new antioxidant, 3,4-dihydroxybiphenyl, with similar ferric reducing capacity to the well-studied piceatannol. The crystal structure of M321A was determined (2.78 Å), and molecular docking of the 3-substituted phenol provided a rational explanation for the altered regioselectivity. Furthermore, HbpA was found to possess pro-S enantioselectivity towards the production of several chiral sulfoxides, whereas variant M321F exhibited improved enantioselectivity. Based on the biochemical characterization of several mutants, it was suggested that Trp97 stabilized the substrate in the active site, Met223 was involved in NADH entrance or binding to the active site, and Pro320 might facilitate FAD movement.

PubMed: 29297973
DOI: 10.1002/cbic.201700648

実験手法

X-RAY DIFFRACTION (2.78 Å)