1887

Abstract

With variously limited chemostat cultures of the glucose consumption rate increased markedly as the concentration of dissolved oxygen (d.o.t.; dissolved oxygen tension) was lowered from 50% to 1 % air saturation. Concomitantly, the specific rate of acetate production increased and lactate, which was not present in the fully aerobic cultures, accumulated in large amounts. Moreover, whereas at a high d.o.t. only an ammonia- limited culture excreted 2-oxoglutarate, all glucose-sufficient cultures excreted this metabolite at a d.o.t. of 1 % air saturation, even more being produced by a K-limited culture than by the ammonia-limited one. The activities of those enzymes of glycolysis that were measured increased in parallel with the glucose consumption rate, as did the activities of enzymes of the Entner-Douderoff pathway. Similarly, the activities of lactate dehydrogenase and acetate kinase (which were synthesized constitutively also) reflected the corresponding metabolite production rates. Tricarboxylic acid (TCA) cycle activity markedly diminished with a lowering of the available oxygen supply and again (with the exception of aconitase and 2-oxoglutarate dehydrogenase) this was mirrored in decreases in the activities of TCA cycle enzymes. Assessments of energy flux in terms of ATP equivalents suggested that it was energetically more expensive to synthesize biomass at a low d.o.t. than at a high one. However, the presence of enzymes of the methylglyoxal bypass (methylglyoxal synthase and glyoxylase) at high activities in cells grown at a low d.o.t. render assessments of ATP flux rates unreliable.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-137-6-1391
1991-06-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/137/6/mic-137-6-1391.html?itemId=/content/journal/micro/10.1099/00221287-137-6-1391&mimeType=html&fmt=ahah

References

  1. Amarasingham C. R., Davis B. D. 1965; Regulation of a-ketoglutarate dehydrogenase formation in Escherichia coli. Journal of Biological Chemistry 240:3664–3670
    [Google Scholar]
  2. Arrigoni C., Singer T. P. 1962; Limitation of the phenazine methosulphate assay for succinic and related dehydrogenases. Nature London: 1931256–1258
    [Google Scholar]
  3. Bergmeyer H. U., Bernt E. 1974; Determination of D-glucose with glucose oxidase and peroxidase. Methods of Enzymatic Analysis, 3. 31205–1215 Bergmeyer H. U. New York & London: Academic Press;
    [Google Scholar]
  4. Bergmeyer H. U., Gawehn K., Grasse M. 1974; Enzymes as biochemical reagents. Methods of Enzymatic Analysis, 3. 1425–522 Bergmeyer H. U. New York & London: Academic Press;
    [Google Scholar]
  5. Bucher T., Pfleiderer G. 1955; Pyruvate kinase from muscle. Methods in Enzymology I:435–440
    [Google Scholar]
  6. Burke R. M., Tempest D. W. 1990; Growth of Bacillus stearothermophilus on glycerol in chemostat culture: expression of an unusual phenotype. Journal of General Microbiology 136:1381–1385
    [Google Scholar]
  7. Cooper R. A. 1984; Metabolism of methylglyoxal in microorganisms. Annual Review of Microbiology 38:49–68
    [Google Scholar]
  8. Cooper R. A., Anderson A. 1970; The formation and catabolism of methylglyoxal during glycolysis. Escherichia coli. FEBS Letters 11:273–276
    [Google Scholar]
  9. Epstein I., Grossowicz N. 1969; Prototrophic thermophilic Bacillus: Isolation, properties and kinetics of growth. Journal of Bacteriology 99:414–417
    [Google Scholar]
  10. Evans C. G. T., Herbert D., Tempest D. W. 1970; The continuous cultivation of micro-organisms. 2. Construction of a chemostat. Methods in Microbiology 2:275–327
    [Google Scholar]
  11. Gibson T., Gordon R. E. 1974; Genus Bacillus.. Bergey's Manual of Determinative Bacteriology, 8.529–550 Buchanan R. E., Gibbons R. E. Baltimore: Williams & Wilkins;
    [Google Scholar]
  12. Harris P., Kornberg H. L. 1972; The uptake of glucose by a thermophilic Bacillus sp. Proceedings of the Royal Society of LondonB182159–170
    [Google Scholar]
  13. Harrison D. E. F. 1976; The regulation of respiration rate in growing bacteria. Advances in Microbial Physiology 14:243–309
    [Google Scholar]
  14. Herbert D„, Phipps P. J., Strange R. E. 1971; Chemical analysis of microbial cells. Methods in Microbiology 5B:209–344
    [Google Scholar]
  15. Hill R. L., Bradshaw R. A. 1969; Fumarase. Methods in Enzymology 13:91–99
    [Google Scholar]
  16. Hillier A. J., Jags G. R. 1982; L-Lactate dehydrogenase, FDP-activated, from Streptococcus cremoris. Methods in Enzymology 89:362–367
    [Google Scholar]
  17. Jones C. W., Brice J. M., Edwards C. 1977; The effect of respiratory chain composition on the growth efficiencies of aerobic bacteria. Archives of Microbiology 115:85–93
    [Google Scholar]
  18. Jones R. G. W., Lascelles J. 1967; The relationship of 4-hydroxybutyric acid to lysine and methionine formation. Escherichia coli. Biochemical Journal 103:709–713
    [Google Scholar]
  19. Kornberg H. L., Reeves R. E. 1972; Correlation between hexose transport and phosphotransferase activity. Escherichia coli. Biochemical Journal 126:1241–1243
    [Google Scholar]
  20. Neijssel O. M., Tempest D. W. 1979; The physiology of metabolic overproduction. Symposia of the Society for General Microbiology 29:53–82
    [Google Scholar]
  21. Nishimura J. S., Griffith M. J. 1981; Acetate kinase from Veillonella alcalescens. Methods in Enzymology 71:311–316
    [Google Scholar]
  22. Noltmann E. A. 1966; Phosphoglucose isomerase. Methods in Enzymology 9:557–568
    [Google Scholar]
  23. Pennock J. 1988 Physiology and energetics of growth of Bacillus stearothermophilus in chemostat culture. PhD thesis University of Sheffield;
    [Google Scholar]
  24. Pennock J., Tempest D. W. 1988; Metabolic and energetic aspects of the growth of Bacillus stearothermophilus in glucose-limited and glucose-sufficient chemostat culture. Archives of Microbiology 150:452–459
    [Google Scholar]
  25. Peterson G. L. 1977; A simplification of the protein assay method of Lowry et al which is more generally applicable. Analytical Biochemistry 83:346–356
    [Google Scholar]
  26. Racker E. 1946; Spectrophotometric measurement of hexokinase and phosphohexokinase activity. Journal of Biological Chemistry 167:843–854
    [Google Scholar]
  27. Reeves H. C., Rabin R., Wegener W. S., Ajl S. J. 1971; Assays of enzymes of the tricarboxylic acid and glyoxylate cycles. Methods in Microbiology6A425–462
    [Google Scholar]
  28. Spiro S., Guest J. R. 1990; FNR and its role in oxygen-regulated gene expression. E. coli. FEMS Microbiology Reviews 75:399–428
    [Google Scholar]
  29. Teixeira de Mattos M. J., Streekstra H., Tempest D. W. 1984; Metabolic uncoupling of substrate-level phosphorylation in anaerobic glucose-limited chemostat cultures of Klebsiella aerogenes NCTC 418. Archives of Microbiology 139:260–264
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-137-6-1391
Loading
/content/journal/micro/10.1099/00221287-137-6-1391
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