pISSN 2288-6982
eISSN 2288-7105




BioDesign 2022; 10(3): 51-55

Published online September 30, 2022

© Korean Society for Structural Biology

Clostridium beijerinckii glyceraldehyde-3-phosphate dehydrogenase GAPDH: purification, crystallization, and X-ray crystallographic analysis

Jie Zhang1,2, Yuanyuan Chen1,2, Tingting Bu1,2, Xue Bai1,2, Shanru He1,2, Lulu Wang1,2, Chunshan Quan1,2 and Yongbin Xu1,2,*

1Department of Bioengineering, College of Life Science, Dalian Minzu University, Dalian 116600, Liaoning, China
2Key Laboratory of Biotechnology and Bioresources Utilization, Ministry of Education, College of Life Science, Dalian Minzu University, Dalian 116600, Liaoning, China

Correspondence to: *

Received: March 16, 2022; Revised: April 13, 2022; Accepted: August 8, 2022


Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a ubiquitous and highly abundant glycolytic enzyme. It plays a pivotal role for the energy and carbon metabolism of most organisms including industrial bacteria. It catalyzes the two step oxidative phosphorylation of D-glyceraldehyde-3-phosphate into 1,3-bisphosphoglycerate using inorganic phosphate and nicotinamide adenine dinucleotide (NAD+) as cofactor. In this study, GAPDH from C. beijerinckii (CbGAPDH) was successfully expressed and purified using Ni-NTA affinity, Q anion-exchange, and gel-filtration chromatography. The protein crystal was obtained and diffracted to a resolution of 1.60 Å. The crystal belonged to the hexagonal space group P6222, with unit-cell parameters of a = 120.6, b = 120.6, and c = 122.1 Å. The Matthews coefficient and solvent content were estimated to be 3.50 Å3 Da–1 and 64.90%, respectively, assuming that the asymmetric unit contained only one recombinant protein molecule.