5B12

Crystal structure of the B-type halohydrin hydrogen-halide-lyase mutant F71W/Q125T/D199H from Corynebacterium sp. N-1074

5B12 の概要

• エントリーDOI: 10.2210/pdb5b12/pdb
• 分子名称: Halohydrin epoxidase B, CHLORIDE ION (3 entities in total)
• 機能のキーワード: lyase, enantioselectivity, halohydrin
• 由来する生物種: Corynebacterium sp.
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 152363.72

構造登録者

Watanabe, F.,Yu, F.,Ohtaki, A.,Yamanaka, Y.,Noguchi, K.,Odaka, M.,Yohda, M. (登録日: 2015-11-17, 公開日: 2016-08-03, 最終更新日: 2024-03-20)

主引用文献

Watanabe, F.,Yu, F.,Ohtaki, A.,Yamanaka, Y.,Noguchi, K.,Odaka, M.,Yohda, M.
Improvement of enantioselectivity of the B-type halohydrin hydrogen-halide-lyase from Corynebacterium sp. N-1074
J.Biosci.Bioeng., 122:270-275, 2016

PubMed Abstract: Halohydrin hydrogen-halide-lyase (H-Lyase) is a bacterial enzyme involved in the degradation of halohydrins. This enzyme catalyzes the intramolecular nucleophilic displacement of a halogen by a vicinal hydroxyl group in halohydrins, producing the corresponding epoxides. The H-Lyases have been classified into A, B and C subtypes based on amino acid sequence similarities. These enzymes have attracted much attention as industrial catalysts in the synthesis of chiral chemicals from prochiral halohydrins. In the present study, we constructed mutants of B-type H-Lyase from Corynebacterium sp. N-1074 (HheB) displaying higher enantioselectivity by structure-based site-directed mutagenesis and random mutagenesis. A triple mutant of HheB exhibited 98.5% enantioselectivity, the highest ever reported, toward (R)-4-chloro-3-hydroxy-butyronitrile production, with the yield reaching approximately two-fold that of the wild-type enzyme. We discuss the structural basis of the high enantioselectivity and productivity of the mutant by comparing the crystal structures of the mutant HheB and the wild-type enzyme in complex with or without the substrate analogue.

PubMed: 27215832
DOI: 10.1016/j.jbiosc.2016.02.003

実験手法

X-RAY DIFFRACTION (1.721 Å)