1887

Abstract

The effect of different carbon sources on the expression of tricarboxylic acid (TCA) cycle genes, along with glyoxylate bypass genes, in was determined. All TCA cycle genes were coordinately expressed in medium containing acetate. Growth in the presence of acetate gave rise to abundant expression of most TCA cycle genes, with the level of transcript being the highest. However, when the cells entered the stationary phase triggered by acetate exhaustion, all genes were repressed, except and , which were slightly induced. Acetate withdrawal from the growth medium during the exponential phase also led to rapid repression of most TCA cycle genes and a corresponding twofold increase in the expression of , which were strongly induced by citrate and succinate. In addition, glucose depletion during the stationary phase led to a corresponding 8–20-fold induction of the , and genes. Addition of glucose to acetate medium resulted in about 10-fold induction of . The strong dependence of TCA cycle and glyoxylate bypass and expression on carbon source availability was confirmed and the regulatory system will be studied precisely.

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2008-10-01
2019-10-18
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References

  1. Arndt, A. & Eikmanns, B. J. ( 2007; ). The alcohol dehydrogenase gene adhA in Corynebacterium glutamicum is subject to carbon catabolite repression. J Bacteriol 189, 7408–7416.[CrossRef]
    [Google Scholar]
  2. Bott, M. ( 2007; ). Offering surprises: TCA cycle regulation in Corynebacterium glutamicum. Trends Microbiol 15, 417–425.[CrossRef]
    [Google Scholar]
  3. Bruckner, R. & Titgemeyer, F. ( 2002; ). Carbon catabolite repression in bacteria: choice of the carbon source and autoregulatory limitation of sugar utilization. FEMS Microbiol Lett 209, 141–148.[CrossRef]
    [Google Scholar]
  4. Brune, I., Werner, H., Huser, A., Kalinowski, J., Puhler, A. & Tauch, A. ( 2006; ). The DtxR protein acting as dual transcriptional regulator directs a global regulatory network involved in iron metabolism of Corynebacterium glutamicum. BMC Genomics 7, 21 [CrossRef]
    [Google Scholar]
  5. Bustin, S. A. ( 2002; ). Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. J Mol Endocrinol 29, 23–39.[CrossRef]
    [Google Scholar]
  6. Cox, D. P. & Hanson, R. S. ( 1968; ). Catabolite repression of aconitate hydratase in Bacillus subtilis. Biochim Biophys Acta 158, 36–44.[CrossRef]
    [Google Scholar]
  7. Cramer, A., Gerstmeir, R., Schaffer, S., Bott, M. & Eikmanns, B. J. ( 2006; ). Identification of RamA, a novel LuxR-type transcriptional regulator of genes involved in acetate metabolism of Corynebacterium glutamicum. J Bacteriol 188, 2554–2567.[CrossRef]
    [Google Scholar]
  8. Cunningham, L., Gruer, M. J. & Guest, J. R. ( 1997; ). Transcriptional regulation of the aconitase genes (acnA and acnB) of Escherichia coli. Microbiology 143, 3795–3805.[CrossRef]
    [Google Scholar]
  9. Dover, L. G., Cerdeno-Tarraga, A. M., Pallen, M. J., Parkhill, J. & Besra, G. S. ( 2004; ). Comparative cell wall core biosynthesis in the mycolated pathogens, Mycobacterium tuberculosis and Corynebacterium diphtheriae. FEMS Microbiol Rev 28, 225–250.[CrossRef]
    [Google Scholar]
  10. Eikmanns, B. J., Thum-Schmitz, N., Eggeling, L., Ludtke, K. U. & Sahm, H. ( 1994; ). Nucleotide sequence, expression and transcriptional analysis of the Corynebacterium glutamicum gltA gene encoding citrate synthase. Microbiology 140, 1817–1828.[CrossRef]
    [Google Scholar]
  11. Eikmanns, B. J., Rittmann, D. & Sahm, H. ( 1995; ). Cloning, sequence analysis, expression, and inactivation of the Corynebacterium glutamicum icd gene encoding isocitrate dehydrogenase and biochemical characterization of the enzyme. J Bacteriol 177, 774–782.
    [Google Scholar]
  12. Eraso, P. & Gancedo, J. M. ( 1984; ). Catabolite repression in yeasts is not associated with low levels of cAMP. Eur J Biochem 141, 195–198.[CrossRef]
    [Google Scholar]
  13. Fouet, A. & Sonenshein, A. L. ( 1990; ). A target for carbon source-dependent negative regulation of the citB promoter of Bacillus subtilis. J Bacteriol 172, 835–844.
    [Google Scholar]
  14. Fouet, A., Jin, S. F., Raffel, G. & Sonenshein, A. L. ( 1990; ). Multiple regulatory sites in the Bacillus subtilis citB promoter region. J Bacteriol 172, 5408–5415.
    [Google Scholar]
  15. Funke, G., von Graevenitz, A., Clarridge, J. E., III & Bernard, K. A. ( 1997; ). Clinical microbiology of coryneform bacteria. Clin Microbiol Rev 10, 125–159.
    [Google Scholar]
  16. Genda, T., Nakamatsu, T. & Ozak, H. ( 2003; ). Purification and characterization of malate dehydrogenase from Corynebacterium glutamicum. J Biosci Bioeng 95, 562–566.[CrossRef]
    [Google Scholar]
  17. Genda, T., Watabe, S. & Ozaki, H. ( 2006; ). Purification and characterization of fumarase from Corynebacterium glutamicum. Biosci Biotechnol Biochem 70, 1102–1109.[CrossRef]
    [Google Scholar]
  18. Gerstmeir, R., Wendisch, V. F., Schnicke, S., Ruan, H., Farwick, M., Reinscheid, D. & Eikmanns, B. J. ( 2003; ). Acetate metabolism and its regulation in Corynebacterium glutamicum. J Biotechnol 104, 99–122.[CrossRef]
    [Google Scholar]
  19. Gerstmeir, R., Cramer, A., Dangel, P., Schaffer, S. & Eikmanns, B. J. ( 2004; ). RamB, a novel transcriptional regulator of genes involved in acetate metabolism of Corynebacterium glutamicum. J Bacteriol 186, 2798–2809.[CrossRef]
    [Google Scholar]
  20. Gruer, M. J. & Guest, J. R. ( 1994; ). Two genetically-distinct and differentially-regulated aconitases (AcnA and AcnB) in Escherichia coli. Microbiology 140, 2531–2541.[CrossRef]
    [Google Scholar]
  21. Gruer, M. J., Bradbury, A. J. & Guest, J. R. ( 1997; ). Construction and properties of aconitase mutants of Escherichia coli. Microbiology 143, 1837–1846.[CrossRef]
    [Google Scholar]
  22. Han, S. O., Inui, M. & Yukawa, H. ( 2007; ). Expression of Corynebacterium glutamicum glycolytic genes varies with carbon source and growth phase. Microbiology 153, 2190–2202.[CrossRef]
    [Google Scholar]
  23. Han, S. O., Inui, M. & Yukawa, H. ( 2008; ). Transcription of Corynebacterium glutamicum genes involved in tricarboxylic acid cycle and glyoxylate cycle. J Mol Microbiol BiotechnolFeb 20 [Epub ahead of print]
    [Google Scholar]
  24. Hanson, R. S. & Cox, D. P. ( 1967; ). Effect of different nutritional conditions on the synthesis of tricarboxylic acid cycle enzymes. J Bacteriol 93, 1777–1787.
    [Google Scholar]
  25. Hayashi, M., Mizoguchi, H., Shiraishi, N., Obayashi, M., Nakagawa, S., Imai, J., Watanabe, S., Ota, T. & Ikeda, M. ( 2002; ). Transcriptome analysis of acetate metabolism in Corynebacterium glutamicum using a newly developed metabolic array. Biosci Biotechnol Biochem 66, 1337–1344.[CrossRef]
    [Google Scholar]
  26. Jin, S. & Sonenshein, A. L. ( 1994; ). Transcriptional regulation of Bacillus subtilis citrate synthase genes. J Bacteriol 176, 4680–4690.
    [Google Scholar]
  27. Kalinowski, J., Bathe, B., Bartels, D., Bischoff, N., Bott, M., Burkovski, A., Dusch, N., Eggeling, L., Eikmanns, B. J. & other authors ( 2003; ). The complete Corynebacterium glutamicum ATCC 13032 genome sequence and its impact on the production of l-aspartate-derived amino acids and vitamins. J Biotechnol 104, 5–25.[CrossRef]
    [Google Scholar]
  28. Kim, H. J., Kim, T. H., Kim, Y. & Lee, H. S. ( 2004; ). Identification and characterization of glxR, a gene involved in regulation of glyoxylate bypass in Corynebacterium glutamicum. J Bacteriol 186, 3453–3460.[CrossRef]
    [Google Scholar]
  29. Kinoshita, S. & Tanaka, K. ( 1972; ). Glutamic acid. In The Microbial Production of Amino Acids, pp. 263–324. Edited by K. Yamada. New York: John Wiley.
  30. Kinoshita, S., Udaka, S. & Shimono, M. ( 1957; ). Studies on the amino acid fermentation Part I. Production of l-glutamic acid by various microorganisms. J Gen Microbiol 3, 193–205.[CrossRef]
    [Google Scholar]
  31. Kolb, A., Busby, S., Buc, H., Garges, S. & Adhya, S. ( 1993; ). Transcriptional regulation by cAMP and its receptor protein. Annu Rev Biochem 62, 749–795.[CrossRef]
    [Google Scholar]
  32. Kotrbova-Kozak, A., Kotrba, P., Inui, M., Sajdok, J. & Yukawa, H. ( 2007; ). Transcriptionally regulated adhA gene encodes alcohol dehydrogenase required for ethanol and n-propanol utilization in Corynebacterium glutamicum R. Appl Microbiol Biotechnol 76, 1347–1356.[CrossRef]
    [Google Scholar]
  33. Kramer, R., Lambert, C., Hoischen, C. & Ebbighausen, H. ( 1990; ). Uptake of glutamate in Corynebacterium glutamicum. 1. Kinetic properties and regulation by internal pH and potassium. Eur J Biochem 194, 929–935.[CrossRef]
    [Google Scholar]
  34. Kromer, J. O., Sorgenfrei, O., Klopprogge, K., Heinzle, E. & Wittmann, C. ( 2004; ). In-depth profiling of lysine-producing Corynebacterium glutamicum by combined analysis of the transcriptome, metabolome, and fluxome. J Bacteriol 186, 1769–1784.[CrossRef]
    [Google Scholar]
  35. Kronemeyer, W., Peekhaus, N., Kramer, R., Sahm, H. & Eggeling, L. ( 1995; ). Structure of the gluABCD cluster encoding the glutamate uptake system of Corynebacterium glutamicum. J Bacteriol 177, 1152–1158.
    [Google Scholar]
  36. Krug, A., Wendisch, V. F. & Bott, M. ( 2005; ). Identification of AcnR, a TetR-type repressor of the aconitase gene acn in Corynebacterium glutamicum. J Biol Chem 280, 585–595.[CrossRef]
    [Google Scholar]
  37. Laemmli, U. K. ( 1970; ). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.[CrossRef]
    [Google Scholar]
  38. Letek, M., Valbuena, N., Ramos, A., Ordonez, E., Gil, J. A. & Mateos, L. M. ( 2006; ). Characterization and use of catabolite-repressed promoters from gluconate genes in Corynebacterium glutamicum. J Bacteriol 188, 409–423.[CrossRef]
    [Google Scholar]
  39. Liebl, W. ( 1991; ). The genus Corynebacterium – nonmedical. In The Prokaryotes, pp. 1157–1171. Edited by A. Balows, H. G. Trüper, M. Dworkin, W. Harder & K. H. Schleifer. New York: Springer-Verlag.
  40. Liebl, W. ( 2005; ). Corynebacterium taxonomy. In Handbook on Corynebacterium glutamicum, pp. 9–34. Edited by L. Eggeling & M. Bott. Boca Raton, FL: CRC Press.
  41. Liu, X. & De Wulf, P. ( 2004; ). Probing the ArcA-P modulon of Escherichia coli by whole genome transcriptional analysis and sequence recognition profiling. J Biol Chem 279, 12588–12597.[CrossRef]
    [Google Scholar]
  42. Lynch, A. S. & Lin, E. C. C. ( 1996; ). Responses to molecular oxygen. In Escherichia coli and Salmonella: Cellular and Molecular Biology, 2nd edn, pp. 1526–1538. Edited by F. C. Neidhardt and others. Washington, DC: American Society for Microbiology.
  43. Molenaar, D., van der Rest, M. E. & Petrovic, S. ( 1998; ). Biochemical and genetic characterization of the membrane-associated malate dehydrogenase (acceptor) from Corynebacterium glutamicum. Eur J Biochem 254, 395–403.[CrossRef]
    [Google Scholar]
  44. Molenaar, D., van der Rest, M. E., Drysch, A. & Yucel, R. ( 2000; ). Functions of the membrane-associated and cytoplasmic malate dehydrogenases in the citric acid cycle of Corynebacterium glutamicum. J Bacteriol 182, 6884–6891.[CrossRef]
    [Google Scholar]
  45. Muffler, A., Bettermann, S., Haushalter, M., Horlein, A., Neveling, U., Schramm, M. & Sorgenfrei, O. ( 2002; ). Genome-wide transcription profiling of Corynebacterium glutamicum after heat shock and during growth on acetate and glucose. J Biotechnol 98, 255–268.[CrossRef]
    [Google Scholar]
  46. Nakano, M. M., Zuber, P. & Sonenshein, A. L. ( 1998; ). Anaerobic regulation of Bacillus subtilis Krebs cycle genes. J Bacteriol 180, 3304–3311.
    [Google Scholar]
  47. Nishimura, T., Vertès, A., Shinoda, Y., Inui, M. & Yukawa, H. ( 2007; ). Anaerobic growth of Corynebacterium glutamicum using nitrate as a terminal electron acceptor. Appl Microbiol Biotechnol 75, 889–897.[CrossRef]
    [Google Scholar]
  48. Ohne, M. ( 1974; ). Regulation of aconitase synthesis in Bacillus subtilis: induction, feedback repression, and catabolite repression. J Bacteriol 117, 1295–1305.
    [Google Scholar]
  49. Perrenoud, A. & Sauer, U. ( 2005; ). Impact of global transcriptional regulation by ArcA, ArcB, Cra, Crp, Cya, Fnr, and Mlc on glucose catabolism in Escherichia coli. J Bacteriol 187, 3171–3179.[CrossRef]
    [Google Scholar]
  50. Przybyla-Zawislak, B., Gadde, D. M., Ducharme, K. & McCammon, M. T. ( 1999; ). Genetic and biochemical interactions involving tricarboxylic acid cycle (TCA) function using a collection of mutants defective in all TCA cycle genes. Genetics 152, 153–166.
    [Google Scholar]
  51. Reinscheid, D. J., Eikmanns, B. J. & Sahm, H. ( 1994a; ). Malate synthase from Corynebacterium glutamicum: sequence analysis of the gene and biochemical characterization of the enzyme. Microbiology 140, 3099–3108.[CrossRef]
    [Google Scholar]
  52. Reinscheid, D. J., Eikmanns, B. J. & Sahm, H. ( 1994b; ). Characterization of the isocitrate lyase gene from Corynebacterium glutamicum and biochemical analysis of the enzyme. J Bacteriol 176, 3474–3483.
    [Google Scholar]
  53. Rosenkrantz, M. S., Dingman, D. W. & Sonenshein, A. L. ( 1985; ). Bacillus subtilis citB gene is regulated synergistically by glucose and glutamine. J Bacteriol 164, 155–164.
    [Google Scholar]
  54. Saier, M. H., Jr & Ramseier, T. M. ( 1996; ). The catabolite repressor/activator (Cra) protein of enteric bacteria. J Bacteriol 178, 3411–3417.
    [Google Scholar]
  55. Saier, M. H., Jr, Chauvaux, S., Cook, G. M., Deutscher, J., Paulsen, I. T., Reizer, J. & Ye, J. J. ( 1996; ). Catabolite repression and inducer control in Gram-positive bacteria. Microbiology 142, 217–230.[CrossRef]
    [Google Scholar]
  56. Unden, G. & Schirawski, J. ( 1997; ). The oxygen-responsive transcriptional regulator FNR of Escherichia coli: the search for signals and reactions. Mol Microbiol 25, 205–210.[CrossRef]
    [Google Scholar]
  57. Unden, G., Achebach, S., Holighaus, G., Tran, H. G., Wackwitz, B. & Zeuner, Y. ( 2002; ). Control of FNR function of Escherichia coli by O2 and reducing conditions. J Mol Microbiol Biotechnol 4, 263–268.
    [Google Scholar]
  58. Usuda, Y., Tujimoto, N., Abe, C., Asakura, Y., Kimura, E., Kawahara, Y., Kurahashi, O. & Matsui, H. ( 1996; ). Molecular cloning of the Corynebacterium glutamicum (Brevibacterium lactofermentum AJ12036) odhA gene encoding a novel type of 2-oxoglutarate dehydrogenase. Microbiology 142, 3347–3354.[CrossRef]
    [Google Scholar]
  59. Wendisch, V. F., de Graaf, A. A., Sahm, H. & Eikmanns, B. J. ( 2000; ). Quantitative determination of metabolic fluxes during coutilization of two carbon sources: comparative analyses with Corynebacterium glutamicum during growth on acetate and/or glucose. J Bacteriol 182, 3088–3096.[CrossRef]
    [Google Scholar]
  60. Wennerhold, J., Krug, A. & Bott, M. ( 2005; ). The AraC-type regulator RipA represses aconitase and other iron proteins from Corynebacterium under iron limitation and is itself repressed by DtxR. J Biol Chem 280, 40500–40508.[CrossRef]
    [Google Scholar]
  61. Winer, J., Jung, C. K. S., Shackel, I. & Williams, P. M. ( 1999; ). Development and validation of real-time quantitative reverse transcriptase-polymerase chain reaction for monitoring gene expression in cardiac myocytes in vitro. Anal Biochem 270, 41–49.[CrossRef]
    [Google Scholar]
  62. Wittmann, C. & De Graaf, A. A. ( 2005; ). Metabolic flux analysis in Corynebacterium glutamicum. In Handbook on Corynebacterium glutamicum, pp. 277–304. Edited by L. Eggeling & M. Bott. Boca Raton, FL: CRC Press.
  63. Yukawa, H., Omumasaba, C. A., Nonaka, H., Kós, P., Okai, N., Suzuki, N., Suda, M., Tsuge, Y., Watanabe, J. & other authors ( 2007; ). Comparative analysis of the Corynebacterium glutamicum group and complete genome sequence of strain R. Microbiology 153, 1042–1058.[CrossRef]
    [Google Scholar]
  64. Zhao, Y. & Lin, Y.-H. ( 2002; ). Flux distribution and partitioning in Corynebacterium glutamicum grown at different specific growth rates. Process Biochem 37, 775–785.[CrossRef]
    [Google Scholar]
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