8HRS

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

Summary for 8HRS

• Entry DOI: 10.2210/pdb8hrs/pdb
• Descriptor: Glyceraldehyde-3-phosphate dehydrogenase, NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE, GLYCEROL, ... (4 entities in total)
• Functional Keywords: dehydrogenase, structural protein
• Biological source: Corynebacterium glutamicum ATCC 13032
• Total number of polymer chains: 4
• Total formula weight: 151848.01

Authors

Son, H.F.,Kim, K.J. (deposition date: 2022-12-15, release date: 2023-12-06)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (2 Å)