8WWS

Crystal structure of cis-epoxysuccinate hydrolase from Klebsiella oxytoca with L(+)-tartaric acid

8WWS の概要

• エントリーDOI: 10.2210/pdb8wws/pdb
• 分子名称: (S)-2-haloacid dehalogenase, L(+)-TARTARIC ACID, 1,2-ETHANEDIOL, ... (6 entities in total)
• 機能のキーワード: cis-epoxysuccinate hydrolase, hydrolase
• 由来する生物種: Klebsiella oxytoca
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 124017.37

構造登録者

Han, Y.,Kong, X.D.,Li, J.,Xu, J.H. (登録日: 2023-10-26, 公開日: 2024-06-12, 最終更新日: 2024-07-03)

主引用文献

Han, Y.,Luo, Y.,Ma, B.D.,Li, J.,Xu, J.H.,Kong, X.D.
Structural Insights of a cis -Epoxysuccinate Hydrolase Facilitate the Development of Robust Biocatalysts for the Production of l-(+)-Tartrate.
Biochemistry, 63:1578-1587, 2024

PubMed Abstract: l-(+)-Tartaric acid plays important roles in various industries, including pharmaceuticals, foods, and chemicals. -Epoxysuccinate hydrolases (CESHs) are crucial for converting -epoxysuccinate to l-(+)-tartrate in the industrial production process. There is, however, a lack of detailed structural and mechanistic information on CESHs, limiting the discovery and engineering of these industrially relevant enzymes. In this study, we report the crystal structures of CESH and CESH-l-(+)-tartrate complex. These structures reveal the key amino acids of the active pocket and the catalytic triad residues and elucidate a dynamic catalytic process involving conformational changes of the active site. Leveraging the structural insights, we identified a robust CESH (550 ± 20 U·mg) with sustained catalytic activity even at a 3 M substrate concentration. After six batches of transformation, immobilized cells with overexpressed CESH maintained 69% of their initial activity, affording an overall productivity of 200 g/L/h. These results provide valuable insights into the development of high-efficiency CESHs and the optimization of biotransformation processes for industrial uses.

PubMed: 38803051
DOI: 10.1021/acs.biochem.4c00141

実験手法

X-RAY DIFFRACTION (1.79 Å)