6S32

Crystal structure of ene-reductase CtOYE from Chroococcidiopsis thermalis.

6S32 の概要

• エントリーDOI: 10.2210/pdb6s32/pdb
• 分子名称: NADH:flavin oxidoreductase/NADH oxidase, ACETATE ION, BENZAMIDINE, ... (5 entities in total)
• 機能のキーワード: ene-reductase, old yellow enzyme, biocatalysis, flavoprotein
• 由来する生物種: Chroococcidiopsis thermalis
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 172695.12

構造登録者

Robescu, M.R.,Niero, M.,Hall, M.,Bergantino, E.,Cendron, L. (登録日: 2019-06-24, 公開日: 2020-01-29, 最終更新日: 2024-01-24)

主引用文献

Robescu, M.S.,Niero, M.,Hall, M.,Cendron, L.,Bergantino, E.
Two new ene-reductases from photosynthetic extremophiles enlarge the panel of old yellow enzymes: CtOYE and GsOYE.
Appl.Microbiol.Biotechnol., 104:2051-2066, 2020

PubMed Abstract: Looking for new ene-reductases with uncovered features beneficial for biotechnological applications, by mining genomes of photosynthetic extremophile organisms, we identified two new Old Yellow Enzyme homologues: CtOYE, deriving from the cyanobacterium Chroococcidiopsis thermalis, and GsOYE, from the alga Galdieria sulphuraria. Both enzymes were produced and purified with very good yields and displayed catalytic activity on a broad substrate spectrum by reducing α,β-unsaturated ketones, aldehydes, maleimides and nitroalkenes with good to excellent stereoselectivity. Both enzymes prefer NADPH but demonstrate a good acceptance of NADH as cofactor. CtOYE and GsOYE represent robust biocatalysts showing high thermostability, a wide range of pH optimum and good co-solvent tolerance. High resolution X-ray crystal structures of both enzymes have been determined, revealing conserved features of the classical OYE subfamily as well as unique properties, such as a very long loop entering the active site or an additional C-terminal alpha helix in GsOYE. Not surprisingly, the active site of CtOYE and GsOYE structures revealed high affinity toward anions caught from the mother liquor and trapped in the anion hole where electron-withdrawing groups such as carbonyl group are engaged. Ligands (para-hydroxybenzaldehyde and 2-methyl-cyclopenten-1-one) added on purpose to study complexes of GsOYE were detected in the enzyme catalytic cavity, stacking on top of the FMN cofactor, and support the key role of conserved residues and FMN cofactor in the catalysis.

PubMed: 31930452
DOI: 10.1007/s00253-019-10287-2

実験手法

X-RAY DIFFRACTION (1.35 Å)