title: Characterization of shikimate dehydrogenase homologues of Corynebacterium glutamicum.
abstract: The function of three Corynebacterium glutamicum shikimate dehydrogenase homologues, designated as qsuD (cgR_0495), cgR_1216, and aroE (cgR_1677), was investigated. A disruptant of aroE required shikimate for growth, whereas a qsuD-deficient strain did not grow in medium supplemented with either quinate or shikimate as sole carbon sources. There was no discernible difference in growth rate between wild-type and a cgR_1216-deficient strain. Enzymatic assays showed that AroE both reduced 3-dehydroshikimate, using NADPH as cofactor, and oxidized shikimate, the reverse reaction, using NADP(+) as cofactor. The reduction reaction was ten times faster than the oxidation. QsuD reduced 3-dehydroquinate using NADH and oxidized quinate using NAD(+) as cofactor. Different from the other two homologues, the product of cgR_1216 displayed considerably lower enzyme activity for both the reduction and the oxidation. The catalytic reaction of QsuD and AroE was highly susceptible to pH. Furthermore, reduction of 3-dehydroshikimate by AroE was inhibited by high concentrations of shikimate, but neither quinate nor aromatic amino acids had any effect on the reaction. Expression of qsuD mRNA was strongly enhanced in the presence of shikimate, whereas that of cgR_1216 and aroE decreased. We conclude that while AroE is the main catalyst for shikimate production in the shikimate pathway, QsuD is essential for quinate/shikimate utilization.
authors.affiliation: Research Institute of Innovative Technology for the Earth, 9-2, Kizugawadai, Kizugawa, Kyoto, 619-0292, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kubota
authors.forename: Yuya
authors.initials: Y
authors.lastname: Tanaka
authors.forename: Kazumi
authors.initials: K
authors.lastname: Hiraga
authors.forename: Masayuki
authors.initials: M
authors.lastname: Inui
authors.forename: Hideaki
authors.initials: H
authors.lastname: Yukawa
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D048230:Corynebacterium glutamicum / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D004795:Enzyme Stability; D007700:Kinetics; D011801:Quinic Acid / Q000378:metabolism; D012765:Shikimic Acid / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D011801:Quinic Acid; D012765:Shikimic Acid; D000429:Alcohol Oxidoreductases; C022641:Shikimate dehydrogenase