7F8D

Malate Dehydrogenase from Geobacillus stearothermophilus (gs-MDH) G218Y mutant

7F8D の概要

• エントリーDOI: 10.2210/pdb7f8d/pdb
• 分子名称: Malate dehydrogenase, NICOTINAMIDE-ADENINE-DINUCLEOTIDE (3 entities in total)
• 機能のキーワード: malate dehydrogenase, oxidoreductase
• 由来する生物種: Geobacillus stearothermophilus (Bacillus stearothermophilus)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 146807.44

構造登録者

Shimozawa, Y.,Himiyama, T.,Nakamura, T.,Nishiya, Y. (登録日: 2021-07-02, 公開日: 2022-02-23, 最終更新日: 2023-11-29)

主引用文献

Shimozawa, Y.,Himiyama, T.,Nakamura, T.,Nishiya, Y.
Increasing loop flexibility affords low-temperature adaptation of a moderate thermophilic malate dehydrogenase from Geobacillus stearothermophilus.
Protein Eng.Des.Sel., 34:-, 2021

PubMed Abstract: Malate dehydrogenase (MDH) catalyzes the reversible reduction of nicotinamide adenine dinucleotide from oxaloacetate to L-malate. MDH from moderate thermophilic Geobacillus stearothermophilus (gs-MDH) has high thermal stability and substrate specificity and is used as a diagnostic reagent. In this study, gs-MDH was engineered to increase its catalytic activity at low temperatures. Based on sequential and structural comparison with lactate dehydrogenase from G. stearothermophilus, we selected G218 as a mutation site to increase the loop flexibility pivotal for MDH catalysis. The G218 mutants showed significantly higher specific activities than the wild type at low temperatures and maintained thermal stability. The crystal structure of the G218Y mutant, which had the highest catalytic efficiency among all the G218 mutants, suggested that the flexibility of the mobile loop was successfully increased by the bulky side chain. Therefore, this study demonstrated the low-temperature adaptation of MDH by facilitating conformational changes during catalysis.

PubMed: 34850194
DOI: 10.1093/protein/gzab026

実験手法

X-RAY DIFFRACTION (2.4 Å)