1887

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). 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 . In this study, the gene, which was annotated as ‘conserved hypothetical protein’ in the genome, was genetically characterized to be essential for growth in minimal medium, and a mutant deleted of was a Phe and Tyr auxotroph. Genetic cloning and expression of in resulted in the formation of a new protein (NCgl0819) having CM activity. It was concluded that encoded the CM of (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---heptulosonate 7-phosphate (DAHP) synthases (DS0950 and DS2098, formerly NCgl0950 and NCgl2098) had been previously identified from . 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 genomes, suggesting a universal occurrence of CM0819-like CMs in the genus .

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.029819-0
2009-10-01
2019-10-15
Loading full text...

Full text loading...

/deliver/fulltext/micro/155/10/3382.html?itemId=/content/journal/micro/10.1099/mic.0.029819-0&mimeType=html&fmt=ahah

References

  1. Ball, S. G., Wickner, R. B., Cottarel, G., Schaus, M. & Tirtiaux, C. ( 1986; ). Molecular cloning and characterization of ARO7–OSM2, a single yeast gene necessary for chorismate mutase activity and growth in hypertonic medium. Mol Gen Genet 205, 326–330.[CrossRef]
    [Google Scholar]
  2. Bradford, M. M. ( 1976; ). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72, 248–254.[CrossRef]
    [Google Scholar]
  3. Calhoun, D. H., Bonner, C. A., Gu, W., Xie, G. & Jensen, R. A. ( 2001; ). The emerging periplasm-localized subclass of AroQ chorismate mutases, exemplified by those from Salmonella typhimurium and Pseudomonas aeruginosa. Genome Biol 2, RESEARCH0030
    [Google Scholar]
  4. Chavez-Bejar, M. I., Lara, A. R., Lopez, H., Hernandez-Chavez, G., Martinez, A., Ramirez, O. T., Bolivar, F. & Gosset, G. ( 2008; ). Metabolic engineering of Escherichia coli for l-tyrosine production by expression of genes coding for the chorismate mutase domain of the native chorismate mutase-prephenate dehydratase and a cyclohexadienyl dehydrogenase from Zymomonas mobilis. Appl Environ Microbiol 74, 3284–3290.[CrossRef]
    [Google Scholar]
  5. Chook, Y. M., Ke, H. & Lipscomb, W. N. ( 1993; ). Crystal structures of the monofunctional chorismate mutase from Bacillus subtilis and its complex with a transition state analog. Proc Natl Acad Sci U S A 90, 8600–8603.[CrossRef]
    [Google Scholar]
  6. Chook, Y. M., Gray, J. V., Ke, H. & Lipscomb, W. N. ( 1994; ). The monofunctional chorismate mutase from Bacillus subtilis. Structure determination of chorismate mutase and its complexes with a transition state analog and prephenate, and implications for the mechanism of the enzymatic reaction. J Mol Biol 240, 476–500.[CrossRef]
    [Google Scholar]
  7. Davidson, B. E. & Hudson, G. S. ( 1987; ). Chorismate mutase-prephenate dehydrogenase from Escherichia coli. Methods Enzymol 142, 440–450.
    [Google Scholar]
  8. Eberhard, J., Raesecke, H. R., Schmid, J. & Amrhein, N. ( 1993; ). Cloning and expression in yeast of a higher plant chorismate mutase. Molecular cloning, sequencing of the cDNA and characterization of the Arabidopsis thaliana enzyme expressed in yeast. FEBS Lett 334, 233–236.[CrossRef]
    [Google Scholar]
  9. Gosset, G., Bonner, C. A. & Jensen, R. A. ( 2001; ). Microbial origin of plant-type 2-keto-3-deoxy-d-arabino-heptulosonate 7-phosphate synthases, exemplified by the chorismate- and tryptophan-regulated enzyme from Xanthomonas campestris. J Bacteriol 183, 4061–4070.[CrossRef]
    [Google Scholar]
  10. Gray, J. V., Golinelli-Pimpaneau, B. & Knowles, J. R. ( 1990; ). Monofunctional chorismate mutase from Bacillus subtilis: purification of the protein, molecular cloning of the gene, and overexpression of the gene product in Escherichia coli. Biochemistry 29, 376–383.[CrossRef]
    [Google Scholar]
  11. Helmstaedt, K., Krappmann, S. & Braus, G. H. ( 2001; ). Allosteric regulation of catalytic activity: Escherichia coli aspartate transcarbamoylase versus yeast chorismate mutase. Microbiol Mol Biol Rev 65, 404–421.[CrossRef]
    [Google Scholar]
  12. Helmstaedt, K., Heinrich, G., Merkl, R. & Braus, G. H. ( 2004; ). Chorismate mutase of Thermus thermophilus is a monofunctional AroH class enzyme inhibited by tyrosine. Arch Microbiol 181, 195–203.[CrossRef]
    [Google Scholar]
  13. Horton, R. M., Hunt, H. D., Ho, S. N., Pullen, J. K. & Pease, L. R. ( 1989; ). Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene 77, 61–68.[CrossRef]
    [Google Scholar]
  14. Hudson, G. S., Wong, V. & Davidson, B. E. ( 1984; ). Chorismate mutase/prephenate dehydrogenase from Escherichia coli K12: purification, characterization, and identification of a reactive cysteine. Biochemistry 23, 6240–6249.[CrossRef]
    [Google Scholar]
  15. Ikeda, M. ( 2005; ). l-Tryptophan production. In Handbook of Corynebacterium glutamicum, pp. 489–51. Edited by L. Eggeling & M. Bott. Boca Raton, FL: CRC Press.
  16. Ikeda, M. ( 2006; ). Towards bacterial strains overproducing l-tryptophan and other aromatics by metabolic engineering. Appl Microbiol Biotechnol 69, 615–626.[CrossRef]
    [Google Scholar]
  17. Ikeda, M. & Nakagawa, S. ( 2003; ). The Corynebacterium glutamicum genome: features and impacts on biotechnological processes. Appl Microbiol Biotechnol 62, 99–109.[CrossRef]
    [Google Scholar]
  18. Jakoby, M., Ngougto-Nkili, C.-E. & Burkovski, A. ( 1999; ). Construction and application of new Corynebacterium glutamicum vectors. Biotechnol Tech 13, 437–441.[CrossRef]
    [Google Scholar]
  19. Jensen, R. A. & Nester, E. W. ( 1965; ). The regulatory significance of intermediary metabolites: control of aromatic acid biosynthesis by feedback inhibition in Bacillus subtilis. J Mol Biol 12, 468–481.[CrossRef]
    [Google Scholar]
  20. Kim, S. K., Reddy, S. K., Nelson, B. C., Vasquez, G. B., Davis, A., Howard, A. J., Patterson, S., Gilliland, G. L., Ladner, J. E. & Reddy, P. T. ( 2006; ). Biochemical and structural characterization of the secreted chorismate mutase (Rv1885c) from Mycobacterium tuberculosis H37Rv: an *AroQ enzyme not regulated by the aromatic amino acids. J Bacteriol 188, 8638–8648.[CrossRef]
    [Google Scholar]
  21. Konopka, A. ( 1993; ). Isolation and characterization of subsurface bacterium that degrades aniline and methylanilines. FEMS Microbiol Lett 111, 93–100.[CrossRef]
    [Google Scholar]
  22. Krappmann, S., Helmstaedt, K., Gerstberger, T., Eckert, S., Hoffmann, B., Hoppert, M., Schnappauf, G. & Braus, G. H. ( 1999; ). The aroC gene of Aspergillus nidulans codes for a monofunctional, allosterically regulated chorismate mutase. J Biol Chem 274, 22275–22282.[CrossRef]
    [Google Scholar]
  23. Krappmann, S., Pries, R., Gellissen, G., Hiller, M. & Braus, G. H. ( 2000; ). HARO7 encodes chorismate mutase of the methylotrophic yeast Hansenula polymorpha and is derepressed upon methanol utilization. J Bacteriol 182, 4188–4197.[CrossRef]
    [Google Scholar]
  24. Kumar, S., Tamura, K. & Nei, M. ( 2004; ). mega3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5, 150–163.[CrossRef]
    [Google Scholar]
  25. Lee, A. Y., Karplus, P. A., Ganem, B. & Vlardy, J. ( 1995; ). Atomic structure of the buried catalytic pocket of Escherichia coli chorismate mutase. J Am Chem Soc 117, 3627–3628.[CrossRef]
    [Google Scholar]
  26. Liebl, W. ( 2005; ). Corynebacterium taxonomy. In Handbook of Corynebacterium glutamicum, pp. 9–36. Edited by L. Eggeling & M. Bott. Boca Raton, FL: CRC Press.
  27. Liu, Y. J., Li, P. P., Zhao, K. X., Wang, B. J., Jiang, C. Y., Drake, H. L. & Liu, S. J. ( 2008; ). Corynebacterium glutamicum contains 3-deoxy-d-arabino-heptulosonate 7-phosphate synthases that display novel biochemical features. Appl Environ Microbiol 74, 5497–5503.[CrossRef]
    [Google Scholar]
  28. MacBeath, G., Kast, P. & Hilvert, D. ( 1998; ). A small, thermostable, and monofunctional chorismate mutase from the archaeon Methanococcus jannaschii. Biochemistry 37, 10062–10073.[CrossRef]
    [Google Scholar]
  29. Prakash, P., Aruna, B., Sardesai, A. A. & Hasnain, S. E. ( 2005; ). Purified recombinant hypothetical protein coded by open reading frame Rv1885c of Mycobacterium tuberculosis exhibits a monofunctional AroQ class of periplasmic chorismate mutase activity. J Biol Chem 280, 19641–19648.[CrossRef]
    [Google Scholar]
  30. Qamra, R., Prakash, P., Aruna, B., Hasnain, S. E. & Mande, S. C. ( 2006; ). The 2.15 Å crystal structure of Mycobacterium tuberculosis chorismate mutase reveals an unexpected gene duplication and suggests a role in host-pathogen interactions. Biochemistry 45, 6997–7005.[CrossRef]
    [Google Scholar]
  31. Sambrook, J., Fritsch, E. F. & Maniatis, T. ( 1989; ). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Sping Harbor Laboratory.
  32. Schafer, A., Tauch, A., Jager, W., Kalinowski, J., Thierbach, G. & Puhler, A. ( 1994; ). Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum. Gene 145, 69–73.[CrossRef]
    [Google Scholar]
  33. Schmidheini, T., Sperisen, P., Paravicini, G., Hutter, R. & Braus, G. ( 1989; ). A single point mutation results in a constitutively activated and feedback-resistant chorismate mutase of Saccharomyces cerevisiae. J Bacteriol 171, 1245–1253.
    [Google Scholar]
  34. Schneider, C. Z., Parish, T., Basso, L. A. & Santos, D. S. ( 2008; ). The two chorismate mutases from both Mycobacterium tuberculosis and Mycobacterium smegmatis: biochemical analysis and limited regulation of promoter activity by aromatic amino acids. J Bacteriol 190, 122–134.[CrossRef]
    [Google Scholar]
  35. Schoner, R. & Herrmann, K. M. ( 1976; ). 3-Deoxy-d-arabino-heptulosonate 7-phosphate synthase. Purification, properties, and kinetics of the tyrosine-sensitive isoenzyme from Escherichia coli. J Biol Chem 251, 5440–5447.
    [Google Scholar]
  36. Shen, X. H., Jiang, C. Y., Huang, Y., Liu, Z. P. & Liu, S. J. ( 2005; ). Functional identification of novel genes involved in the glutathione-independent gentisate pathway in Corynebacterium glutamicum. Appl Environ Microbiol 71, 3442–3452.[CrossRef]
    [Google Scholar]
  37. Shiio, I. & Sugimoto, S. ( 1979; ). Two components of chorismate mutase in Brevibacterium flavum. J Biochem 86, 17–25.
    [Google Scholar]
  38. Siehl, D. L. ( 1997; ). Inhibitors of EPSP synthase, glutamine synthetase and histidine synthesis. In Herbicide Activity: Toxicology, Biochemistry and Molecular Biology, pp. 37–67. Edited by R. M. Roe, J. D. Burton & R. J. Kurh. Amsterdam/Oxford: IOS.
  39. Strater, N., Schnappauf, G., Braus, G. & Lipscomb, W. N. ( 1997; ). Mechanisms of catalysis and allosteric regulation of yeast chorismate mutase from crystal structures. Structure 5, 1437–1452.[CrossRef]
    [Google Scholar]
  40. Sugimoto, S. & Shiio, I. ( 1980a; ). Purification and properties of dissociable chorismate mutase from Brevibacterium flavum. J Biochem 88, 167–176.
    [Google Scholar]
  41. Sugimoto, S. & Shiio, I. ( 1980b; ). Purification and properties of bifunctional 3-deoxy-d-arabino-heptulosonate 7-phosphate synthetase-chorismate mutase component A from Brevibacterium flavum. J Biochem 87, 881–890.
    [Google Scholar]
  42. Tauch, A., Kassing, F., Kalinowski, J. & Puhler, A. ( 1995; ). The Corynebacterium xerosis composite transposon Tn5432 consists of two identical insertion sequences, designated IS1249, flanking the erythromycin resistance gene ermCX. Plasmid 34, 119–131.[CrossRef]
    [Google Scholar]
  43. Tauch, A., Kirchner, O., Loffler, B., Gotker, S., Puhler, A. & Kalinowski, J. ( 2002; ). Efficient electrotransformation of Corynebacterium diphtheriae with a mini-replicon derived from the Corynebacterium glutamicum plasmid pGA1. Curr Microbiol 45, 362–367.[CrossRef]
    [Google Scholar]
  44. Yakandawala, N., Romeo, T., Friesen, A. D. & Madhyastha, S. ( 2008; ). Metabolic engineering of Escherichia coli to enhance phenylalanine production. Appl Microbiol Biotechnol 78, 283–291.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.029819-0
Loading
/content/journal/micro/10.1099/mic.0.029819-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error