1887

Microbiology Society logo

Volume 142, Issue 12

Metabolic adaptation of to growth rate and glucose availability Free

Abstract

The parasitic protist adapted the specific activities of twelve of the enzymes involved in glucose metabolism to the growth rate and glucose availability. These changes in enzyme activities were induced by culturing in chemostats with glucose, present in rate-limiting or excess concentrations, as carbon and energy source. The specific activities were measured in pelleted cells at each steady state, while metabolic end products were determined in filtered culture fluid. The specific activities were lower in cells grown on growth-rate-limiting concentrations of glucose and higher in organisms cultured in the presence of excess glucose. In both cases enzyme activities were higher at increasing growth rates. For most enzymes the difference between the highest and lowest activities was an order of magnitude. The specific activities of eleven of the enzymes were strongly correlated to each other (correlation coefficients 0·83-0·99), the exception being lactate dehydrogenase. The rates of production of the three major end products, lactate, acetate and glycerol, increased with increasing growth rates. Alanine was not formed in measurable quantities. The ratio of the end products formed was strongly influenced by the growth rates and glucose availability. The rates of formation of acetate and glycerol correlated best with the specific activities of the enzymes catalysing the final reactions of their respective pathways. This suggests that the production of acetate and glycerol is rate-limited by these final steps. In contrast, the formation of lactate did not correlate with the specific activity of lactate dehydrogenase but was determined by the rate of glucose consumption.

  • Published Online:
Keyword(s): chemostat , glucose , Keywords: energy metabolism , metabolic control analysis and Trichomonas vaginalis
© Society for General Microbiology 1996
Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-142-12-3337
1996-12-01
2024-04-16
Loading full text...

Full text loading...

/deliver/fulltext/micro/142/12/mic-142-12-3337.html?itemId=/content/journal/micro/10.1099/13500872-142-12-3337&mimeType=html&fmt=ahah

References

  1. Arese P., Cappuccinelli P. 1974; Glycolysis and pentose phosphate cycle in Trichomonas vaginalis. I. Enzyme activity pattern and the constant proportion quintet. Int J Biochem 5:859–865
     [Google Scholar]
  2. Avilan L., Garcia P. 1994; Hysteresis of cytosolic NADP-malic enzyme I1 from Trypanosoma crqi. Mol Biocbem Parasitol 65:225–232
     [Google Scholar]
  3. Bergmeyer H. U. 1974; Enzymes as biochemical reagents: hexokinase. In Methods of Enzymatic Analysis= pp. 473 Edited by Bergmeyer H. U. New York: Academic Press;
     [Google Scholar]
  4. 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
     [Google Scholar]
  5. Cazzulo I. J., Franke de Cazzulo B. M., Engel I. C., Cannata J. J. 1985; End products and enzyme levels of aerobic glucose fermentation in trypanosomatids. Mol Biocbem Parasitol l6:329–343
     [Google Scholar]
  6. Chapman A., Linstead D. J., Lloyd D., Williams J. 1985; 13C- NMR reveals glycerol as an unexpected major metabolite of the protozoan parasite Trichomonas vaginalis. FEBS Eett 191:287–292
     [Google Scholar]
  7. Coombs G. H., Müller M. 1995; Energy metabolism in anaerobic protozoa. In Biochemistry and Molecular Biology of Parasites pp. 33–47 Edited by Marr J. J., Müller M. New York: Academic Press;
     [Google Scholar]
  8. Davies S. E., Brindle K. M. 1992; Effects of overexpression of phosphofructokinase on glycolysis in the yeast Saccharomyces cerevisiae. Biochemistry 31:4729–4735
     [Google Scholar]
  9. Diamond L. S. 1957; The establishment of various trichomonads of animals and man in axenic cultures. J Parasitol 43:488–490
     [Google Scholar]
  10. Eggstein M., Kuhlmann E. 1974; Triglycerides and glycerol: determination after alkaline hydrolysis. In Methods of Enzymatic Analysis pp. 1825–1835 Edited by Bergmeyer H. U. Weinheim: Verlag Chemie;
     [Google Scholar]
  11. Fouts A. C., Kraus S. J. 1980; Trichomonas vaginalis: reevaluation of its clinical presentation and laboratory diagnosis. J Infect Dis 141:137–143
     [Google Scholar]
  12. Gutmann I., Wahlefeld A. W. 1974; L-( + )-Lactate: deter¬mination with lactate dehydrogenase and NAD. In Methods of Enzymatic Analysis pp. 1464–1475 Edited by Bergmeyer H. U. Weinheim: Verlag Chemie;
     [Google Scholar]
  13. Hochachka P. W., Somero G. N. 1984 Biochemical Adaptation, 2nd edn.. Princeton: Princeton University Press;
     [Google Scholar]
  14. Hrdy I., Mertens E., Van Schaftingen E. 1993; Identification, purification and separation of different isozymes of NADP-specific malic enzyme from Tritrichomonas foetus. Mol Biochem Parasitol 57:253–260
     [Google Scholar]
  15. Kacser H., Burns J. A., Fell D. A. 1995; The control of flux. Biochem Soc Trans 23:341–366
     [Google Scholar]
  16. Lehker M. W., Alderete J. F. 1990; Properties of Trichomonas vaginalis grown under chemostat controlled growth conditions. Genitourin Med 66:193–199
     [Google Scholar]
  17. Lloyd D., Kristensen B. 1985; Metronidazole inhibition of hydrogen production in vivo in drug-sensitive and resistant strains of Trichomonas vaginalis. J Gen Microbiol 131:849–853
     [Google Scholar]
  18. Lloyd D., Paget T. A. 1991; The effects of environmental factors on the metabolism of Giardia and Trichomonas. In Biochemical Protozoology pp. 92–101 Edited by Coombs G. H., North M. J. London: Taylor & Francis;
     [Google Scholar]
  19. Mack S. R., Müller M. 1978; Effect of oxygen and carbon dioxide on the growth of Trichomonas vaginalis and Tritrichomonas foetus. J Parasitol 64:927–929
     [Google Scholar]
  20. Mack S. R., Müller M. 1980; End products of carbohydrate metabolism in Trichomonas vaginalis. Comp Biochem Physiol 67B:213–216
     [Google Scholar]
  21. Markos A., Miretsky A., Müller M. 1993; A glyceraldehyde-3- phosphate dehydrogenase with eubacterial features in the amito- chondriate eukaryote, Trichomonas vaginalis. J Mol Evol 37:631–643
     [Google Scholar]
  22. Mertens E. 1991; Pyrophosphate-dependent phosphofructo¬kinase, an anaerobic glycolytic enzyme?. FEBS Lett 285:1–5
     [Google Scholar]
  23. Mertens E. 1993; ATP versus pyrophosphate: glycolysis revisited in parasitic protists. Parasitol Today 9:122–126
     [Google Scholar]
  24. Mertens E., Müller M. 1990; Glucokinase and fructokinase of Trichomonas vaginalis and Tritrichomonas foetus. J Protocol 37:384–388
     [Google Scholar]
  25. Mertens E., Van Schaftingen E., Müller M. 1989; Presence of a fructose-2,6-bisphosphate-insensitive pyrophosphate: fructose-6- phosphate phosphotransferase in the anaerobic protozoa Tritrichomonas foetus, Trichomonas vaginalis and Isotricha prostoma. Mol Biochem Parasitol 37:183–190
     [Google Scholar]
  26. Misset O., Opperdoes F. R. 1984; Simultaneous purification of hexokinase, class-I fructose-bisphosphate aldolase, triosephosphate isomerase and phosphoglycerate kinase from Trypanosoma brucei. Eur J Biochem 144:475–483
     [Google Scholar]
  27. Misset O., Van Beeumen J., Lambeir A. M., Van der Meer R., Opperdoes F. R. 1987; Glyceraldehyde-phosphate dehydrogen¬ase from Trypanosoma brucei. Comparison of the glycosomal and cytosolic isoenzymes. Eur J Biochem 162:501–507
     [Google Scholar]
  28. Müller M. 1988; Energy metabolism of protozoa without mitochondria. Annu Rev Microbiol 42:465–488
     [Google Scholar]
  29. Müller M. 1989; Biochemistry of Trichomonas vaginalis. In Trichomonads Parasitic in Humans pp. 53–83 Edited by Honigberg B. M. New York: Springer;
     [Google Scholar]
  30. Müller M. 1991; Energy metabolism of anaerobic parasitic protists. In Biochemical Protozoology pp. 80–91 Edited by Coombs G. H., North M. J. London: Taylor & Francis;
     [Google Scholar]
  31. Müller S„, Boles E., May M., Zimmermann F. K. 1995; Different internal metabolites trigger the induction of glycolytic gene expression in Saccharomyces cerevisiae. J Bacteriol 177:4517–4519
     [Google Scholar]
  32. Paget T. A., Lloyd D. 1990; Trichomonas vaginalis requires traces of oxygen and high concentrations of carbon dioxide for optimal growth. Mol Biochem Parasitol 41:65–72
     [Google Scholar]
  33. Schaaff I., Heinisch J., Zimmermann F. K. 1989; Overproduction of glycolytic enzymes in yeast. Yeast 5:285–290
     [Google Scholar]
  34. Sierkstra L. N., ter Schure E. G., Verbakel J. M. A., Verrips C. T. 1994; A nitrogen-limited, glucose-repressed, continuous culture of Saccharomyces cerevisiae. Microbiology 140:593–599
     [Google Scholar]
  35. Steinbüchel A., Müller M. 1986a; Anaerobic pyruvate metabolism of Tritrichomonas foetus and Trichomonas vaginalis hydro- genosomes. Mol Biochem Parasitol 20:57–65
     [Google Scholar]
  36. Steinbüchel A., Müller M. 1986b; Glycerol, a metabolic end product of Trichomonas vaginalis and Tritrichomonas foetus. Mol Biochem Parasitol 20:45–55
     [Google Scholar]
  37. Ter Kuile B. H. 1994a; Carbohydrate metabolism and physiology of the parasitic protist Trichomonas vaginalis studied in chemostats. Microbiology 140:2495–2502
     [Google Scholar]
  38. Ter Kuile B. H. 1994b; Adaptation of the carbon metabolism of Trichomonas vaginalis to the nature and availability of the carbon source. Microbiology 140:2503–2510
     [Google Scholar]
  39. Ter Kuile B. H., Opperdoes F. R. 1991; Chemostat cultures of Eeishmania donovani promastigotes and Trypanosoma brucei procyclic trypomastigotes. Mol Biochem Parasitol 45:171–173
     [Google Scholar]
  40. Thauer R. K., Jungermann K., Decker K. 1977; Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Reviews 41:100–180
     [Google Scholar]
  41. Williamson D. H. 1974; L-Alanine: determination with alanine dehydrogenase. In Methods of Enzymatic Analysis pp. 1679–1682 Edited by Bergmeyer H. U. New York: Academic Press;
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-142-12-3337
Loading
Metabolic adaptation of Trichomonas vaginalis to growth rate and glucose availability
Microbiology 142, 3337 (1996); https://doi.org/10.1099/13500872-142-12-3337
/content/journal/micro/10.1099/13500872-142-12-3337
/content/journal/micro/10.1099/13500872-142-12-3337
Loading

Data & Media loading...

Most cited Most Cited RSS feed

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