Article info Vol. 5  No. 1   pp.  35 ~ 39
Title Purification, crystallization and X-ray crystallographic analysis of cystathionine gamma-synthase from Corynebacterium glutamicum
Authors Hye-Young Sagong and Kyung-Jin Kim*
Institutions School of Life Sciences, KNU Creative BioResearch Group, Kyungpook National University, Daehak-ro 80, Buk-ku, Daegu 41566, Korea. *Correspondence: kkim@knu.ac.kr
Abstract Cystathionine gamma synthase from Corynebacterium glutamicum (CgMetB) is a key enzyme for the production of L-methionine and it condenses O-acetyl-L-homoserine (OAHS) and cysteine to produce cystathionine. MetB is also an attractive target for the development of antimicrobial compounds because it catalyzes the first reaction of the L-methionine biosynthetic pathway. The CgMetB was overexpressed and purified to homogeneity by affinity and size-exclusion chromatography. The CgMetB protein was crystallized using hanging-drop vapor-diffusion method in the presence of 13% polyethylene glycol 3350 and 0.1 M Magnesium formate dihydrate at 295 K. X-ray diffraction data were collected to a maximum resolution of 1.5 Å. The crystal belonged to space group F222, with unit cell parameters a = 58.57 Å, b = 149.85 Å, c = 161.86, α = β = γ = 90.0°. With one molecules per asymmetric unit, the crystal volume per unit protein mass was 2.13 Å 3 Da -1 , which correspond to a solvent content of approximately 42.27%.