1887

Abstract

Periplasmic oxidation of glucose into gluconate and 2-ketogluconate in occurs via glucose dehydrogenase (GDH) and gluconate dehydrogenase (GaDH), respectively. Since, as is shown here, in the presence of glucose, gluconate and 2-ketogluconate are not further metabolized intracellularly the physiological function of this periplasmic route was studied. It was found that periplasmic oxidation of glucose could function as an alternative production route of ATP equivalents. Instantaneous activation of either GDH or GaDH reduced the rate of degradation of glucose via glycolysis and the tricarboxylic acid (TCA) cycle . Furthermore, aerobic, magnesium- and phosphate-limited chemostat cultures with glucose as the carbon source showed high GDH plus GaDH activities in contrast to nitrogen-and sulphate-limited cultures. However, when fructose, which is not degraded by GDH, was the carbon source, specific oxygen consumption rates under these four conditions were essentially the same. The latter observation suggests that high transmembrane phosphate gradients which are supposedly present under phosphate-limited conditions do not cause high energetic demands due to futile cycling of phosphate ions. In addition, dissipation of the transmembrane phosphate gradient of phosphate-limited cells immediately increased the rate of intracellular glucose degradation. It is concluded that under phosphate-limited conditions (i) extensive futile cycling of phosphate ions is absent and (ii) low concentrations of phosphate ions limit intracellular degradation of glucose. Glyceraldehyde-3-phosphate dehydrogenase (GADPH) activities of cell-free extracts of glucose-grown cells harvested from aerobic chemostat cultures limited in various nutrients showed that at least a tenfold overcapacity in GAPDH activity was present under phosphate-limited conditions with respect to the steady-state carbon fluxes through this enzyme. The physiological significance of this adaptation and the possible role of GDH and GaDH are discussed.

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1994-09-01
2021-07-27
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References

  1. Aiking H., Stijnman A., Van Garderen C., Van Heerikhuizen H., Vant Riet J. Inorganic phosphate accumulation and cadmium detoxification in Klebsiella aerogenes NCTC 418 growing in continuous culture. Appl Environ Microbiol 1984; 47:374–377
    [Google Scholar]
  2. Ameyama M., Nonobe M., Shinagawa E., Matsushita K., Takimoto K., Adachi O. Purification and characterization of the quinoprotein D-glucose dehydrogenase apoenzyme from Escherichia coli. Agric Biol Chem 1986; 50:49–57
    [Google Scholar]
  3. Beardmore-Gray M., Anthony C. The oxidation of glucose In Acinetobacter calcoaceticus: interaction of the quinoprotein glucose dehydrogenase with the electron transport chain. J Gen Microbiol 1986; 132:1257–1268
    [Google Scholar]
  4. Bergmeyer H.U., Gawehn K., Grassl M. Enzyme als biochemische Reagentien. In Methoden der engymatischen Analyse 1974 Edited by Bergmeyer H.U. Weinheim Germany: Verlag Ghemie; 1 pp 454–557
    [Google Scholar]
  5. Buurman E.T., Boiardi J., L, Teixeira De Mattos M.J., Neijssel O.M. The role of magnesium and calcium ions in the glucose dehydrogenase activity of Klebsiella pneumoniae NCTC 418. Arch Microbiol 1990; 153:502–505
    [Google Scholar]
  6. Buurman E.T., Teixeira De Mattos M.J., Neijssel O.M. Futile cycling of potassium and ammonium ions in Escherichia coli. In Alcali Cation Transport in Procaryotes 1992 Edited by Bakker E.P. Boca Raton, FL: CRC Press; pp 411–426
    [Google Scholar]
  7. Cooper R.A. Metabolism of methylglyoxal in micro-organisms. Ann Rev Microbiol 1984; 38:49–68
    [Google Scholar]
  8. Evans C.G.T., Herbert D., Tempest D.W. The continuous cultivation of micro-organisms II Construction of a chemostat. In Methods in Microbiology 1970 Edited by Norris J.R., Ribbons D.W. London and New York: Academic Press; 2 pp 277–327
    [Google Scholar]
  9. Fraenkel D.G. Glycolysis, pentose-phosphate pathway, and Entner-Doudoroff pathway. In Escherichia coli and Salmonella typhimurium 1987 Edited by Neidhardt F.C. Washington, DC: American Society for Microbiology; pp 142–150
    [Google Scholar]
  10. Gornall A.G., Bardawill C.J., David M.A. Determination of serum proteins by means of the biuret reaction. J Biol Chem 1949; 177:751–766
    [Google Scholar]
  11. Hardy G.P.M.A., Teixeira De Mattos M.J., Neijssel O.M. Energy conservation by pyrroloquinoline quinone-linked xylose oxidation in Pseudomonasputida NCTC 10936 during carbon-limited growth in chemostat culture. FEMS Microbiol Lett 1993; 107:107–110
    [Google Scholar]
  12. Herbert D. Stoicheiometric aspects of microbial growth. In Continuous Culture. 6. Applications and New Fields 1976 Edited by Dean A.C.R., Ellwood D.C., Evans C.G.T., Melling J. Chichester: Ellis Horwood; pp 1–30
    [Google Scholar]
  13. Herbert D., Phipps P.J., Strange R.E. Chemical analysis of microbial cells. Methods in Microbiology 1971; 5B:209–344
    [Google Scholar]
  14. Hommes R.W.J., Van Hell B., Postma P.W., Neijssel O.M., Tempest D.W. The functional significance of glucose dehydrogenase in Klebsiella aerogenes. Arch Microbiol 1985; 143:163–168
    [Google Scholar]
  15. Hommes R.W.J., Herman P.T.D., Postma P.W., Tempest D.W., Neijssel O.M. The separate roles of PQQ and apoenzyme syntheses in the regulation of glucose dehydrogenase activity in Klebsiella pneumoniae NCTC 418. Arch Microbiol 1989a; 151:257–260
    [Google Scholar]
  16. Hommes R.W.J., Postma P.W., Neijssel O.M., Tempest D.W. The influence of culture pH value on the direct glucose oxidative pathway in Klebsiella pneumoniae NCTC 418. Arch Microbiol 1989b; 151:261–267
    [Google Scholar]
  17. Hopper D.J., Cooper R.A. The regulation of Escherichia coli methylglyoxal synthase; A new control site in glycolysis. FEBS Lett 1971; 13:213–216
    [Google Scholar]
  18. Matsushita K., Shinagawa E., Ameyama M. D-Gluconate dehydrogenase from bacteria, 2-keto-D-gluconate-vielding, membrane bound. Methods Enzymol 1982; 89:187–193
    [Google Scholar]
  19. Matsushita K., Nonobe M., Shinagawa E., Adachi O., Ameyama M. Reconstitution of a pyrroloquinoline quinone-dependent D-glucose oxidase respiratory chain of Escherichia coli with cytochrome o-oxidase. J Bacterial 1987; 169:205–209
    [Google Scholar]
  20. Meury J., Kepes A. The regulation of potassium fluxes for the adjustment and maintenance of potassium levels in Escherichia coli. Eur J Biochem 1981; 119:165–170
    [Google Scholar]
  21. Mulder M.M., Teixeira De Mattos M.J., Postma P.W., Van Dam K. Energetic consequences of multiple K+ uptake systems. Biochim Biophys Acta 1986; 851:223–228
    [Google Scholar]
  22. Neijssel O.M. The effect of 2,4-dinitrophenol on the growth of Klebsiella aerogenes NCTC 418 in aerobic chemostat cultures. FEA4S Microbiol Lett 1977; 1:47–50
    [Google Scholar]
  23. Neijssel O.M., Buurman E.T., Teixeira De Mattos M.J. The role of futile cycles in the energetics of bacterial growth. Biochim Biophys Acta 1990; 1018:252–255
    [Google Scholar]
  24. Neijssel O.M., Tempest D.W. Production of gluconic acid and 2-ketogluconic acid by Klebsiella aerogenes. Arch Microbiol 1975; 105:183–185
    [Google Scholar]
  25. Neijssel O.M., Tempest D.W., Postma P.W., Duine J.A., Frank Jzn J. Glucose metabolism by KMimited Klebsiella aerogenes: evidence for the involvement of a quinoprotein glucose dehydrogenase. EEMS Microbiol Lett 1983; 20:35–39
    [Google Scholar]
  26. Nelson D.L., Kennedy E.P. Transport of magnesium by a repressible and a non-repressible system in Escherichia coli. Proc Natl Acad Sci USA 1972; 69:1091–1093
    [Google Scholar]
  27. Peterson G.L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem 1977; 83:346–356
    [Google Scholar]
  28. Rao N.N., Torriani A. Molecular aspects of phosphate transport in Escherichia coli. Mol Microbiol 1990; 4:1083–1090
    [Google Scholar]
  29. Rosenberg H., Gerdes R.G., Chegwidden K. Two systems for the uptake of phosphate in Escherichia coli. J Bacteriol 1977; 131:505–511
    [Google Scholar]
  30. Stewart V. Nitrate respiration in relation to facultative metabolism in Enterobacteria. Microbiol Rev 1988; 52:190–232
    [Google Scholar]
  31. Streekstra H. Metabolic uncoupling in anaerobic Klebsiella pneumoniae 1990 PhD thesis, University of Amsterdam, The Netherlands;
    [Google Scholar]
  32. Van Schie B.J., Hellingwerf K.J., Van Dijken J.P., Elferink M.G.L., Van Dijl J.M., Kuenen J.G., Konings W.N. Energy transduction by electron transfer via a pvrroloquinoline quinone-dependent glucose dehydrogenase in Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter calcoaceticus (var Iwoffi). J Bacteriol 1985; 136:493–499
    [Google Scholar]
  33. Van Schie B.J., Rouwenhorst R.J., De Bont J.A.M., Van Dijken J.P., Kuenen J.G. An in vivo analysis of the energetics of aldose oxidation by Acinetobacter calcoaceticus. Appl Microbiol Biotechnol 1987; 26:560–567
    [Google Scholar]
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