Open AccessOverexpression, crystallization, and preliminary X‐ray crystallographic analysis of shikimate dehydrogenase from Thermotoga maritima
Author(s) -
Lee Hyung Ho
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111019877
Subject(s) - thermotoga maritima , orthorhombic crystal system , shikimate pathway , escherichia coli , crystallization , crystallography , ammonium sulfate , chemistry , monomer , stereochemistry , biochemistry , crystal structure , gene , biosynthesis , organic chemistry , polymer
Shikimate dehydrogenase (SDH), which catalyses the NADPH‐dependent reduction of 3‐dehydroshikimate to shikimate in the shikimate pathway, is an attractive target for the development of herbicides and antimicrobial agents. Previous structural studies showed that SDH exists in two conformations, an open form and a closed form, and it is believed that the conformational state is crucial to understanding a catalytic mechanism. To facilitate further structural comparisons among SDHs, structural analysis of an SDH from Thermotoga maritima encoded by the Tm0346 gene has been initiated. SDH from T. maritima has been overexpressed in Escherichia coli and crystallized at 296 K using ammonium sulfate as a precipitant. Crystals of T. maritima SDH diffracted to 1.45 Å resolution and belonged to orthorhombic space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 54.21, b = 62.45 and c = 68.68 Å. The asymmetric unit contains a monomer, with a corresponding V M of 2.01 Å 3 Da −1 and a solvent content of 38.9% by volume.