8HRS

Crystal structure of glyceraldehyde-3-phosphate dehydrogenase from Corynebacterium glutamicum ATCC13032 (L36S/T37K/P192S) in complex with NADP

8HRS の概要

• エントリーDOI: 10.2210/pdb8hrs/pdb
• 分子名称: Glyceraldehyde-3-phosphate dehydrogenase, NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE, GLYCEROL, ... (4 entities in total)
• 機能のキーワード: dehydrogenase, structural protein
• 由来する生物種: Corynebacterium glutamicum ATCC 13032
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 151848.01

構造登録者

Son, H.F.,Kim, K.J. (登録日: 2022-12-15, 公開日: 2023-12-06)

主引用文献

Son, H.F.,Yu, H.,Hong, J.,Lee, D.,Kim, I.K.,Kim, K.J.
Structure-Guided Protein Engineering of Glyceraldehyde-3-phosphate Dehydrogenase from Corynebacterium glutamicum for Dual NAD/NADP Cofactor Specificity.
J.Agric.Food Chem., 71:17852-17859, 2023

PubMed Abstract: Since the discovery of l-glutamate-producing , it has evolved to be an industrial workhorse. For biobased chemical production, suppling sufficient amounts of the NADPH cofactor is crucial. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a glycolytic enzyme that converts glyceraldehyde-3-phosphate (G3P) to 1,3-bisphosphoglycerate and produces NADH, is a major prospective solution for the cofactor imbalance issue. In this study, we determined the crystal structure of GAPDH from ATCC13032 (GAPDH). Based on the structural information, we generated six GAPDH variants, GAPDH, GAPDH, GAPDH, GAPDH, GAPDH, and GAPDH, that can produce both NADH and NAPDH. The final GAPDH variant showed a 212-fold increase in enzyme activity for NADP as well as 200% and 30% increased activity for the G3P substrate under NAD and NADP cofactor conditions, respectively. In addition, crystal structures of GAPDH variants in complex with NAD(P) permit the elucidation of differences between wild-type GAPDH and variants in relation to cofactor stabilization.

PubMed: 37935620
DOI: 10.1021/acs.jafc.3c06176

実験手法

X-RAY DIFFRACTION (2 Å)