4A3U

X-structure of the old yellow enzyme homologue from zymomonas mobilis (NCR)

4A3U の概要

• エントリーDOI: 10.2210/pdb4a3u/pdb
• 分子名称: NADH\:FLAVIN OXIDOREDUCTASE/NADH OXIDASE, FLAVIN MONONUCLEOTIDE, ACETATE ION, ... (7 entities in total)
• 機能のキーワード: oxidoreductase
• 由来する生物種: ZYMOMONAS MOBILIS
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 80493.47

構造登録者

Hoeffken, H.W. (登録日: 2011-10-04, 公開日: 2012-10-17, 最終更新日: 2023-12-20)

主引用文献

Reich, S.,Hoeffken, H.W.,Rosche, B.,Nestl, B.M.,Hauer, B.
Crystal Structure Determination and Mutagenesis Analysis of the Ene Reductase Ncr.
Chembiochem, 13:2400-, 2012

PubMed Abstract: The crystal structure of the "ene" nicotinamide-dependent cyclohexenone reductase (NCR) from Zymomonas mobilis (PDB ID: 4A3U) has been determined in complex with acetate ion, FMN, and nicotinamide, to a resolution of 1.95 Å. To study the activity and enantioselectivity of this enzyme in the bioreduction of activated α,β-unsaturated alkenes, the rational design methods site- and loop-directed mutagenesis were applied. Based on a multiple sequence alignment of various members of the Old Yellow Enzyme family, eight single-residue variants were generated and investigated in asymmetric bioreduction. Furthermore, a structural alignment of various ene reductases predicted four surface loop regions that are located near the entrance of the active site. Four NCR loop variants, derived from loop-swapping experiments with OYE1 from Saccharomyces pastorianus, were analysed for bioreduction. The three enzyme variants, P245Q, D337Y and F314Y, displayed increased activity compared to wild-type NCR towards the set of substrates tested. The active-site mutation Y177A demonstrated a clear influence on the enantioselectivity. The loop-swapping variants retained reduction efficiency, but demonstrated decreased enzyme activity compared with the wild-type NCR ene reductase enzyme.

PubMed: 23033175
DOI: 10.1002/CBIC.201200404

実験手法

X-RAY DIFFRACTION (1.7 Å)