Open AccessExpression, purification, crystallization and preliminary crystallographic analysis of Cg1458: a novel oxaloacetate decarboxylase from Corynebacterium glutamicum
Author(s) -
Ran Tingting,
Wang Yu,
Xu Dongqing,
Wang Weiwu
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111023220
Subject(s) - corynebacterium glutamicum , decarboxylation , citrate synthase , crystallization , orthorhombic crystal system , chemistry , stereochemistry , crystallography , nuclear chemistry , biochemistry , crystal structure , organic chemistry , enzyme , catalysis , gene
Oxaloacetate decarboxylase catalyses the decarboxylation of oxaloacetate to pyruvate and CO 2 . Recently, the Corynebacterium glutamicum gene product Cg1458 was determined to be a soluble oxaloacetate decarboxylase. To elucidate the mechanism of oxaloacetate decarboxylation by Cg1458, recombinant Cg1458 was purified and crystallized. The best crystal was grown from 0.2 M MgCl 2 , 0.1 M Bis‐Tris pH 6.0, 25%( w / v ) polyethylene glycol 3350 using the hanging‐drop method. The crystals belonged to space group P 4 3 2 1 2, with unit‐cell parameters a = b = 124.1, c = 73.6 Å. The crystals are most likely to contain a dimer in the asymmetric unit, with a V M value of 2.27 Å 3 Da −1 . A full data set was collected at 1.9 Å resolution using synchrotron radiation on beamline BL17U of SSRF, Shanghai, China. Structure‐solution attempts by molecular replacement were successful with PDB entries 3qdf or 2dfu as the template.