Continuous flow method for extraction and bioluminescence assay of ATP in baker's yeast

  • Maj-Rita Siro
  • Henrik Romar
  • Timo Lövgren
Article

Summary

The intracellular ATP of baker's yeast (Saccharomyces cerevisiae) was measured using the bioluminescent firefly luciferase assay. Benzalkonium chloride and trichloro-acetic acid served in the experiments as extracting agents and optimal conditions for the extraction and assay of the intracellular ATP are reported. Using the results obtained from manually performed experiments two continuous flow systems were designed for the measurement of ATP in yeast cells during cell growth. Good correlation between the amount of cellular ATP and cell growth was found during the exponential growth phase.

Keywords

Yeast Cell Saccharomyces Cerevisiae Saccharomyces Exponential Growth Continuous Flow 

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References

  1. Alexander DN, Ederer GM, Matsen JN (1976) Evaluation of an adenosine 5′ triphosphate assay as a screening method to detect significant bacteriuria. J Clin Microbiol 3:42–46Google Scholar
  2. Chappelle EW, Levin GV (1968) Use of the firefly bioluminescent reaction for rapid detection and counting of bacteria. Biochem Med 2:41–52Google Scholar
  3. Guinn G, Eidenbock MP (1972) Extraction, purification and estimation of ATP from leaves, floral buds and immature fruits of cotton. Anal Biochem 50:89–97Google Scholar
  4. Holm-Hansen O, Karl DM (1978). Biomass and adenylate energy charge determination in microbial cell extracts and environmental samples. In: DeLuca MA (ed) Methods in enzymology 57. Academic Press, New York London, p 73–85Google Scholar
  5. Holmsen H, Holmsen I, Bernhardsen A (1966) Microdetermination of adenosine diphosphate and adenosine triphosphate in plasma with the firefly luciferase system. Anal Biochem 17:456–473Google Scholar
  6. Klofat W, Picciolo G, Chappelle EW, Freese E (1969) Production of adenosine triphosphate in normal cells and sporulation mutants of Bacillus subtilis. J Biol Chem 244:3270–3276Google Scholar
  7. Levin GV, Usdin E, Slonium AR (1968) Rapid detection of microorganisms in aerospace water systems. Aerosp Med 1:14–16Google Scholar
  8. Lin S, Cohen HP (1968) Measurement of adenosine triphosphate content of crayfish stretch receptor cell preparations. Anal Biochem 24:531–540Google Scholar
  9. Lundin A, Thore A (1975a) Analytical information obtainable by evaluation of the time course of firefly bioluminescence in the assay of ATP. Anal Biochem 66:47–63Google Scholar
  10. Lundin A, Thore A (1975b) Comparison of methods for extraction of bacterial adenine nucleotides determined by firefly assay. Appl Microbiol 30:713–721Google Scholar
  11. Lundin A, Rickardsson, A, Thore A (1976) Continuous monitoring of ATP-converting reactions by purified firefly luciferase. Anal Biochem 75:611–620Google Scholar
  12. Mc Elroy WD (1963) Crystalline firefly luciferase. In: Colowick SP, Kaplan NO (eds) Methods in enzymology VI. Academic Press, New York London, p 445–448Google Scholar
  13. Mc Elroy WD, Green A (1956) Function of adenosine triphosphate in the activation of luciferin. Arch Biochem Biophys 64:257–271Google Scholar
  14. Polakis ES, Bartley W (1966) Changes in the intracellular concentrations of adenosine phosphates and nicotinamide nucleotides during the aerobic growth cycle of yeast on different carbon sources. Biochem J 99:521–533Google Scholar
  15. Rasmussen H, Nielsen R (1968) An improved analysis of adenosine triphosphate by the luciferase method. Acta Chem Scand 22:1745–1756Google Scholar
  16. Scharff TG, Maupin WC (1960) Correlation of the metabolic effects of benzalkonium chloride with its membrane effects in yeast. Biochem Pharmacol 5:79–86Google Scholar
  17. Sharpe AN, Woodrow MN, Jackson AK (1970) Adenosine triphosphate (ATP) levels in foods contaminated by bacteria. J Appl Bacteriol 33:758–767Google Scholar
  18. Siro MR, Lövgren T (1979) Influence of glucose on the α-glucoside permease activity of yeast. J Appl Microbiol Biotechnol 7:59–66Google Scholar
  19. Spielmann H, Jacob-Müller V, Schulz P (1981) Simple assay of 0.1–1.0 pmol of ATP, ADP and AMP in single somatic cells using purified luciferin luciferase. Anal Biochem 113:172–178Google Scholar
  20. StJoiin JB (1970) Determination of ATP in Chlorella with the luciferin luciferase enzyme system. Anal Biochem 37:409–416Google Scholar
  21. Thore A (1979) Technical aspects of the bioluminescent firefly luciferase assay of ATP. Sci Tools 26:30–34Google Scholar
  22. Thore A, Ånsehn S, Lundin A, Bergman S (1975) Detection of bacteriuria by luciferase assay of adenosine triphosphate. J. Clin Microbiol 1:1–8Google Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • Maj-Rita Siro
    • 1
  • Henrik Romar
    • 1
  • Timo Lövgren
    • 2
  1. 1.Department of Biochemistry and PharmacyÅbo AkademiTurku 50Finland
  2. 2.Wallac Biochemical LaboratoryTurku 10Finland

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