4FN4

short-chain NAD(H)-dependent dehydrogenase/reductase from Sulfolobus acidocaldarius

4FN4 の概要

• エントリーDOI: 10.2210/pdb4fn4/pdb
• 分子名称: Short chain dehydrogenase, NICOTINAMIDE-ADENINE-DINUCLEOTIDE, GLYCEROL, ... (5 entities in total)
• 機能のキーワード: nadh-binding, rossmann fold, oxidoreductase
• 由来する生物種: Sulfolobus acidocaldarius
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 110902.53

構造登録者

Pennacchio, A.,Sannino, V.,Sorrentino, G.,Rossi, M.,Raia, C.A.,Esposito, L. (登録日: 2012-06-19, 公開日: 2012-08-15, 最終更新日: 2023-09-13)

主引用文献

Pennacchio, A.,Sannino, V.,Sorrentino, G.,Rossi, M.,Raia, C.A.,Esposito, L.
Biochemical and structural characterization of recombinant short-chain NAD(H)-dependent dehydrogenase/reductase from Sulfolobus acidocaldarius highly enantioselective on diaryl diketone benzil.
Appl.Microbiol.Biotechnol., 97:3949-3964, 2013

PubMed Abstract: The gene encoding a novel alcohol dehydrogenase that belongs to the short-chain dehydrogenases/reductases superfamily was identified in the aerobic thermoacidophilic crenarchaeon Sulfolobus acidocaldarius strain DSM 639. The saadh2 gene was heterologously overexpressed in Escherichia coli, and the resulting protein (SaADH2) was purified to homogeneity and both biochemically and structurally characterized. The crystal structure of the SaADH2 NADH-bound form reveals that the enzyme is a tetramer consisting of identical 27,024-Da subunits, each composed of 255 amino acids. The enzyme has remarkable thermophilicity and thermal stability, displaying activity at temperatures up to 80 °C and a 30-min half-inactivation temperature of ∼88 °C. It also shows good tolerance to common organic solvents and a strict requirement for NAD(H) as the coenzyme. SaADH2 displays a preference for the reduction of alicyclic, bicyclic and aromatic ketones and α-ketoesters, but is poorly active on aliphatic, cyclic and aromatic alcohols, showing no activity on aldehydes. Interestingly, the enzyme catalyses the asymmetric reduction of benzil to (R)-benzoin with both excellent conversion (98 %) and optical purity (98 %) by way of an efficient in situ NADH-recycling system involving a second thermophilic ADH. The crystal structure of the binary complex SaADH2-NADH, determined at 1.75 Å resolution, reveals details of the active site providing hints on the structural basis of the enzyme enantioselectivity.

PubMed: 22805786
DOI: 10.1007/s00253-012-4273-z

実験手法

X-RAY DIFFRACTION (1.75 Å)