Archives of Microbiology

, Volume 135, Issue 1, pp 63–67 | Cite as

Nucleotide pools of growing, synchronized and stressed cultures of Saccharomyces cerevisiae

  • Günther Ditzelmüller
  • Wilfried Wöhrer
  • Christian P. Kubicek
  • Max Röhr
Original Papers


High pressure liquidd chromatography has been used to study the acid soluble nucleotide pool of Saccharomyces cerevisiae under different conditions of growth. ATP, ADP, AMP, NAD, GTP, UTP, UDP, CTP, CDP, and UDP-sugars plus UMP could be separated and were found in concentrations higher than 0.1 μmol per g yeast cell dry weight (=detection limit). During glucose-limited continuous culture the levels of individual nucleotides depended on the growth rate, which was most pronounced with pyrimidine (uridine, cytidine) nucleotides. The energy charge (E.C.) remained high (0.9) at all growth rates (0.07–0.3 h-1). During synchronized growth at a constant growth rate (0.11 h-1) almost all nucleotide levels and the E.C. remained at constant values with the only exception of UDP-sugars and UMP of which increased levels were found during the phase of budding. Under conditions of metabolic stress (addition of antimycin A, deoxyglucose plus iodoacetate) pronounced changes in the levels of purine (adenine and guanine) nucleotides and the E.C. were observed. All other nucleotides were less influenced by these conditions. Only under these conditions IMP accumulation was observed. The results strongly argue against the significance of purine nucleotide or E.C. measurements under viable conditions. In contrast, changes in the levels of pyrimidine nucleotides seem to be indicative of changes in the flux through the metabolic pathways where they act as coenzymes.

Key words

Nucleotide pools Continuous cultivation Synchronized growth Saccharomyces cerevisiae 



High pressure liquid chromatography


  1. Bach HP, Wöhrer W, Röhr M (1978) Continuous determination of ethanol during aerobic cultivation of yeasts. Biotechnol Bioeng 20:799–807Google Scholar
  2. Cabib E, Duran A, Bowers B (1979) Localized activation of chitin synthetase in the initiation of yeast septum formation. In: Burnett JH, Trinci APJ (eds) Fungal walls and hypahl growth. Cambridge University Press, Cambridge, pp 189–202Google Scholar
  3. Chapman AG, Atkinson DE (1977) Adenine nucleotide concentrations and turnover rates. Their correlation with biological activity in bacteria and yeast. Adv Microb Physiol 15:253–306Google Scholar
  4. Chow FK, Grushka E (1979) High performance liquid chromatography of nucleotides and nucleosides using outersphere and innersphere metal-solute complexes. J Chromatogr 185:361–373Google Scholar
  5. Ditzelmüller G, Wöhrer W, Kubicek CP, Röhr M (1981) Erfassung des intracellulären Nucleotidpools von Saccharomyces cerevisiae mittels HPLC. Österr Chem Zeitschr 82:248Google Scholar
  6. Engelberg J (1964) Measurement of degrees of synchrony in cell populations. In: Zeuthen E (ed) Synchrony in cell division and growth. Interscience Publishers, New York, p 497Google Scholar
  7. Fiechter A, von Meyenburg K (1968) Automatic analysis of gas exchange in microbial systems. Biotechnol Bioeng 10:535–540Google Scholar
  8. Harrison DEF, Maitra PK (1969) Control of respiration and metabolism in growing Klebsiella aerogenes. The role of adenine nucleotides. Biochem J 112:647–656Google Scholar
  9. Hoffman NE, Liao JC (1977) Reversed phase high performance liquid chromatographic separations of nucleotides in the presence of solvophobic ions. Anal Chem 49:2231–2234Google Scholar
  10. Karl DM (1980) Cellular nucleotide measurements and applications in microbial ecology. Microbiol Rev 44:739–796Google Scholar
  11. Knowles CJ (1977) Microbial metabolic regulation by adenine nucleotide pools. Symp Soc Gen Microbiol 27:241–283Google Scholar
  12. Kuenzi MT, Fiechter A (1972) Regulation of carbohydrate composition of Saccharomyces cerevisiae under growth limitation. Arch Mikrobiol 84:254–265Google Scholar
  13. Mangat BS (1971) Changes in the free nucleotide pool during growth in cultures of Polytoma uvella. Can J Biochem 49:811–815Google Scholar
  14. Noda L (1977) Adenylate kinase. In: Boyer PD (ed) The enzymes, 3rd edn, vol 8. Academic Press, New York, pp 279–305Google Scholar
  15. Sabina RL, Dalke P, Hanks AR, Magill JM, Magill CW (1981) Changes in nucleotide pools during conidial germination in Neurospora crassa. Can J Biochem 59:899–905Google Scholar
  16. Savioja T, Miettinen JK (1966) Isolation and identification of acid soluble phosphorus compounds of yeast. Acta Chem Scand 20:2435–2443Google Scholar
  17. Slayman CL (1973) Adenine nucleotide levels in Neurospora as influenced by conditions of growth and by metabolic inhibtors. J Bacteriol 114:752–766Google Scholar
  18. Schatzmann H (1974) Anaerobes Wachstum von Saccharomyces cerevisiae. Diss. ETH Zürich No. 5504Google Scholar
  19. Swedes JS, Dial ME, McLaughlin CS (1979) Regulation of protein synthesis during energy limitation of Saccharomyces cerevisiae. J Bacteriol 138:162–170Google Scholar
  20. von Meyenburg K (1969) Katabolit — Repression und der Sprossungszyklus von Saccharomyces cerevisiae. Vierteljahresschr Naturforsch Ges Zürich 114:113–222Google Scholar
  21. Weibel K (1973) Zur Energetik von Saccharomyces cerevisiae. Diss. ETH, No. 5155, ZürichGoogle Scholar
  22. Wöhrer W, Röhr M (1981) Regulatory aspects of bakers' yeast metabolism in aerobic fed-batch cultures. Biotechnol Bioeng 23:567–581Google Scholar
  23. Wöhrer W, Forstenlehner L, Röhr M (1981) Evaluation of the Crabtree effect in different yeasts grown in batch and continuous culture. In: Stewart GG, Russell I (eds) Current development in yeast research. Pergamon Press, Toronto, pp 405–410Google Scholar

Copyright information

© Springer-Verlag 1983

Authors and Affiliations

  • Günther Ditzelmüller
    • 1
  • Wilfried Wöhrer
    • 1
  • Christian P. Kubicek
    • 1
  • Max Röhr
    • 1
  1. 1.Institut für Biochemische Technologie und MikrobiologieTechnische Universität WienWienAustria

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