1887

Abstract

The anaerobic parasitic protist was adapted in chemostats to eight different conditions defined by different growth rates and carbon regimes. Glucose or maltose was used as carbon and energy source. Cells cultured under well-defined steady states were tested in short-term experiments. The kinetics of glucose and maltose uptake were determined and their glucokinase and α-glucosidase activities were measured. Uptake in 20 min was measured with radiolabeled glucose and maltose, rather than analogues, using the silicone oil centrifugation technique. Hence, the accumulated label represents both transport and metabolic activity. The total uptake of glucose was highest in organisms that had been starved for glucose during growth. The kinetics of glucose uptake can be understood by assuming rate-limitation by transport across the plasma membrane at low external concentrations and by the subsequent metabolism at concentrations exceeding a cross-over value. The specific glucokinase activity correlated in only four out of eight cases with the saturation uptake. The kinetics of maltose uptake indicated rate-limitation at low maltose concentrations by a diffusion-limited step and at higher levels by metabolic steps. The uptake of maltose was primarily affected by the growth rate during culture, the highest growth rates resulting in most uptake. Maltose uptake was determined only partially by the cellular α-glucosidase activity. The activities of both transport and metabolic enzymes changed due to the culture conditions suggesting that the control over glucose and maltose metabolism is shared by several steps in the pathway.

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1994-09-01
2024-04-27
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References

  1. Bergmeyer H.U. Enzymes as biochemical reagents: hexokinase. In Methods of Enzymatic Analysis 1974 Edited by Bergmeyer H.U. New York: Academic Press; p 473
    [Google Scholar]
  2. Bisson L.F., Coons D.M., Kruckeberg A.L., Lewis D.A. Yeast sugar transporters. Crit Rev Biochem Mol Biol 1993; 28:259–308
    [Google Scholar]
  3. Colowick S.P. The hexokinases. In The Enzymes 1973 Edited by Boyer P.D. New York: Academic Press; 9 part B pp 1–48
    [Google Scholar]
  4. Diamond L.S. The establishment of various trichomonads of animals and man in axenic cultures. J Parasitol 1957; 43:488–490
    [Google Scholar]
  5. Gasnier B. Characterization of low-and high-affinity glucose transporters in the yeast Kluyveromyces marxianus. Biochim Biopbys Acta 1987; 903:425–433
    [Google Scholar]
  6. Heinrich R., Rapoport S.M. Metabolic regulation and mathematical models. Progr Biophys Mol Biol 1977; 32:1–82
    [Google Scholar]
  7. Hill R., Whittingham C.P. Photosynthesis 1955 London: Methuen;
    [Google Scholar]
  8. Hochachka P.W., Somero G.N. Biochemical Adaptation 1984 Princeton, N J: Princeton University Press;
    [Google Scholar]
  9. Kacser H., Porteous J.W. Control of metabolism: what do we have to measure. Trends Biochem Sci 1987; 12:5–14
    [Google Scholar]
  10. Kaplan A., Badger M.R., Berry J.A. Photosynthesis and the intracellular inorganic carbon pool in the blue-green alga Anabaena variabilis: response to external C02 concentration. Planta 1980; 149:219–266
    [Google Scholar]
  11. Lehker M.W., Alderete J.F. Properties of Trichomonas vaginalis grown under chemostat controlled growth conditions. Genitourin Med 1990; 66:193–199
    [Google Scholar]
  12. Mertens E., Muller M. Glucokinase and fructokinase of Trichomonas vaginalis and Tritrichomonas foetus. J Protocol 1990; 37:384–388
    [Google Scholar]
  13. Muller M. Energy metabolism of anaerobic parasitic protists. In Biochemical Protozoology 1991 Edited by Coombs G.H., North M.J. London: Taylor and Francis; pp 80–91
    [Google Scholar]
  14. Opperdoes F.R. Compartmentation of carbohydrate metabolism in trypanosomes. Annu Rev Microbiol 1987; 41:127–151
    [Google Scholar]
  15. Peinado J.M., Cameira-Dos-Santos P.J., Loureido-Dias M.C. Regulation of glucose transport in Candida utilis. J Gen Microbiol 1988; 134:195–201
    [Google Scholar]
  16. Petterson G., Petterson P. Ultimate limits for the reaction flux and metabolite levels that may be evolutionarily reached in a linear metabolic pathway. Eur J Biochem 1990; 194:135–139
    [Google Scholar]
  17. Postma E., Scheffers W.A., Van Dijken J.P. Adaptation of the kinetics of glucose transport to environmental conditions in the yeast Candida utilis CBS 621: a continuous-culture study. J Gen Microbol 1988; 134:1109–116
    [Google Scholar]
  18. Stein W.D. Transport and Diffusion Across Cell Membranes 1986 London : Academic Press;
    [Google Scholar]
  19. Ter Kuile B.H. Glucose and proline transport in kinetoplastids. Parasitology Today 1993; 9:206–210
    [Google Scholar]
  20. Ter Kuile B.H. Carbohydrate metabolism and physiology of the parasitic protist Trichomonas vaginalis studied in chemostats. Microbiology 1994; 140:2495–2502
    [Google Scholar]
  21. Ter Kuile B.H., Cook M. The kinetics of facilitated diffusion followed by enzymatic conversion of the substrate. Biochim Biophys Acta 1994 (in press);
    [Google Scholar]
  22. Ter Kuile B.H., Muller M. Interaction between facilitated diffusion of glucose across the plasma membrane and its metabolism in Trichomonas vaginalis. FEMS Microbiol Lett 1993; 110:27–32
    [Google Scholar]
  23. Ter Kuile B.H., Muller M. Maltose utilization by extracellular hydrolysis followed by glucose transport in the amitochondriate eukaryote, Trichomonas vaginalis. Parasitology 1994 (in press);
    [Google Scholar]
  24. Ter Kuile B.H., Opperdoes F.R. Glucose uptake by Trypanosoma brucei. Rate-limiting steps in glycolysis and regulation of the glycolytic flux. J Biol Chem 1991; 266:857–862
    [Google Scholar]
  25. Ter Kuile B.H., Opperdoes F.R. Comparative physiology of two protozoan parasites, Leishmania donovani and Trypanosoma brucei, grown in chemostats. J Bacterial 1992a; 174:2929–2934
    [Google Scholar]
  26. Ter Kuile B.H., Opperdoes F.R. A chemostat study on proline uptake and metabolism of Leishmania donovani. J Protocol 1992b; 39:555–558
    [Google Scholar]
  27. Ter Kuile B.H., Opperdoes F.R. Mutual adjustment of glucose uptake and metabolism in Trypanosoma brucei grown in the chemostat. J Bacteriol 1992c; 174:1273–1279
    [Google Scholar]
  28. Ter Kuile B.H., Opperdoes F.R. Uptake and turnover of glucose in Leishmania donovani. Mol Biochem Parasitol 1993; 60:313–322
    [Google Scholar]
  29. Thomson A.B.R., Dietschy J.M. The role of the unstirred water layer in intestinal permeation. In Pharmacology of Intestinal Permeation II 1984 Edited by Csaky T.Z. Berlin: Springer-Verla; pp 165–269
    [Google Scholar]
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