title: Genetic and biochemical identification of the chorismate mutase from Corynebacterium glutamicum.
abstract: Chorismate mutase (CM) catalyses the rearrangement of chorismate to prephenate and is also the first and the key enzyme that diverges the shikimate pathway to either tryptophan (Trp) or phenylalanine (Phe) and tyrosine (Tyr). Corynebacterium glutamicum is one of the most important amino acid producers for the fermentation industry and has been widely investigated. However, the gene(s) encoding CM has not been experimentally identified in C. glutamicum. In this study, the ncgl0819 gene, which was annotated as 'conserved hypothetical protein' in the C. glutamicum genome, was genetically characterized to be essential for growth in minimal medium, and a mutant deleted of ncgl0819 was a Phe and Tyr auxotroph. Genetic cloning and expression of ncgl0819 in Escherichia coli resulted in the formation of a new protein (NCgl0819) having CM activity. It was concluded that ncgl0819 encoded the CM of C. glutamicum (CM0819). CM0819 was demonstrated to be a homodimer and is a new member of the monofunctional CMs of the AroQ structural class. The CM0819 activity was not affected by Phe, Tyr or Trp. Two 3-deoxy-d-arabino-heptulosonate 7-phosphate (DAHP) synthases (DS0950 and DS2098, formerly NCgl0950 and NCgl2098) had been previously identified from C. glutamicum. CM0819 significantly stimulated DAHP synthase (DS2098) activity. Physical interaction between CM0819 and DS2098 was observed. When CM0819 was present, DS2098 activity was subject to allosteric inhibition by chorismate and prephenate. Conserved hypothetical proteins homologous to CM0819 were identified in all known Corynebacterium genomes, suggesting a universal occurrence of CM0819-like CMs in the genus Corynebacterium.
authors: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, PR China.
authors: Pan-Pan
authors: PP
authors: Li
authors: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, PR China.
authors: Ya-Jun
authors: YJ
authors: Liu
authors: Environmental Microbiology and Biotechnology Research Center, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, PR China.|State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, PR China.
authors: Shuang-Jiang
authors: SJ
authors: Liu
mesh_terms: D000494:Allosteric Regulation; D002826:Chorismate Mutase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D002827:Chorismic Acid / Q000378:metabolism; D003001:Cloning, Molecular; D048230:Corynebacterium glutamicum / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D003509:Cyclohexanecarboxylic Acids / Q000378:metabolism; D053138:Cyclohexenes / Q000378:metabolism; D019281:Dimerization; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D020043:Genes, Essential; D010649:Phenylalanine / Q000096:biosynthesis; D010802:Phylogeny; D011485:Protein Binding; D025941:Protein Interaction Mapping; D017386:Sequence Homology, Amino Acid; D014443:Tyrosine / Q000096:biosynthesis
chemical_list: D003509:Cyclohexanecarboxylic Acids; D053138:Cyclohexenes; D014443:Tyrosine; D010649:Phenylalanine; D002826:Chorismate Mutase; D002827:Chorismic Acid; C005550:prephenic acid