5YW4

Structure-Guided Engineering of Reductase: Efficient Attenuating Substrate Inhibition in Asymmetric Catalysis

5YW4 の概要

• エントリーDOI: 10.2210/pdb5yw4/pdb
• 分子名称: Protein induced by osmotic stress, NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE (3 entities in total)
• 機能のキーワード: reductase, nadp+, asymmetric catalysis, oxidoreductase
• 由来する生物種: Scheffersomyces stipitis (Yeast)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 37749.27

構造登録者

Shang, Y.P.,Chen, Q.,Li, A.T.,Yu, H.L.,Xu, J.H. (登録日: 2017-11-28, 公開日: 2019-03-06, 最終更新日: 2023-11-22)

主引用文献

Shang, Y.P.,Chen, Q.,Li, A.T.,Quan, S.,Xu, J.H.,Yu, H.L.
Attenuated substrate inhibition of a haloketone reductase via structure-guided loop engineering.
J.Biotechnol., 308:141-147, 2020

PubMed Abstract: Substrate inhibition of enzymes is one of the main obstacles encountered frequently in industrial biocatalysis. Haloketone reductase SsCR was seriously inhibited by substrate 2,2',4'-trichloroacetophenone. In this study, two essential loops were found that have a relationship with substrate binding by conducting X-ray crystal structure analysis. Three key residues were selected from the tips of the loops and substituted with amino acids with lower hydrophobicity to weaken the hydrophobic interactions that bridge the two loops, resulting in a remarkable reduction of substrate inhibition. Among these variants, L211H showed a significant attenuation of substrate inhibition, with a K of 16 mM, which was 16 times that of the native enzyme. The kinetic parameter k/K of L211H was 3.1 × 10 s mM, showing the comparable catalytic efficiency to that of the wild-type enzyme (WT). At the substrate loading of 100 mM, the space time yield of variant L211H in asymmetric reduction of the haloketone was 3-fold higher than that of the WT.

PubMed: 31866427
DOI: 10.1016/j.jbiotec.2019.12.011

実験手法

X-RAY DIFFRACTION (2.047 Å)