1887

Abstract

The mechanisms by which the weak acid preservative benzoic acid inhibits the growth of have been investigated. A reduction in the pyruvate kinase level, which decreases glycolytic flux, did not increase the sensitivity of yeast to benzoic acid. However, a decrease in 6-phosphofructo-1-kinase (PF1K), which does not affect glycolytic flux, did increase sensitivity to benzoic acid. Also, resistance was increased by elevating PF1K levels. Hence, resistance to benzoic acid was not dependent upon optimum glycolytic flux, but upon an adequate PF1K activity. Benzoic acid was shown to depress fructose 2,6-bisphosphate levels in YKC14, a mutant with low PF1K levels. This effect was partially suppressed by overexpressing constitutively active 6-phosphofructo-2-kinase (Pfk26) or by inactivating fructose-2,6-bisphosphatase (in a Δ mutant). The inactivation of PF2K (in a Δ Δ mutant) increased benzoic acid sensitivity. Therefore, the antimicrobial effects of benzoic acid can be relieved, at least in part, by the genetic manipulation of PF1K or fructose 2,6-bisphosphate levels.

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2001-02-01
2019-12-13
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References

  1. Avigad, G. ( 1981; ). Stimulation of yeast phosphofructokinase activity by fructose 2,6-bisphosphate. Biochem Biophys Res Commun 102, 985-991.[CrossRef]
    [Google Scholar]
  2. Bartrons, R., Van Schaftingen, E., Vissers, S. & Hers, H.-G. ( 1982; ). The stimulation of yeast phosphofructokinase by fructose-2,6-bisphosphate. FEBS Lett 143, 137-140.[CrossRef]
    [Google Scholar]
  3. Boles, E., Göhlmann, H. W. H. & Zimmermann, F. K. ( 1996; ). Cloning of a second gene encoding 6-phosphofructo-2-kinase in yeast, and characterisation of mutant strains without fructose-2,6-bisphosphate. Mol Microbiol 20, 65-76.[CrossRef]
    [Google Scholar]
  4. Booth, I. R. & Kroll, R. G. ( 1989; ). The preservation of foods by low pH. In Mechanisms of Action of Food Preservation Procedures , pp. 119-160. Edited by G. W. Gould. London:Elsevier.
  5. Bradford, M. M. ( 1976; ). A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein-dye binding. Anal Biochem 72, 248-254.[CrossRef]
    [Google Scholar]
  6. Church, G. M. & Gilbert, W. ( 1984; ). Genomic sequencing. Proc Natl Acad Sci USA 81, 1991-1995.[CrossRef]
    [Google Scholar]
  7. Davies, S. E. C. & Brindle, K. M. ( 1992; ). Effects of overexpression of phosphofructokinase on glycolysis in the yeast Saccharomyces cerevisiae. Biochemistry 31, 4729-4735.[CrossRef]
    [Google Scholar]
  8. Eraso, P. & Gancedo, J. M. ( 1987; ). Activation of yeast plasma membrane ATPase by acid pH during growth. FEBS Lett 224, 187-192.[CrossRef]
    [Google Scholar]
  9. Feinberg, A. P. & Vogelstein, B. ( 1983; ). A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132, 6-13.[CrossRef]
    [Google Scholar]
  10. François, J., Van Schaftingen, E. & Hers, H.-G. ( 1986; ). Effect of benzoate on the metabolism of fructose-2,6-bisphosphate in yeast. Eur J Biochem 154, 141-145.[CrossRef]
    [Google Scholar]
  11. François, J., Van Schaftingen, E. & Hers, H.-G. ( 1988; ). Characterisation of phosphofructokinase 2 and of enzymes involved in the degradation of fructose-2,6-bisphosphate in yeast. Eur J Biochem 171, 599-608.[CrossRef]
    [Google Scholar]
  12. Gietz, R. D. & Woods, R. A. ( 1998; ). Transformation of yeast by the lithium acetate/single-stranded carrier DNA/PEG method. In Yeast Gene Analysis: Methods in Microbiology , pp. 53-66. Edited by A. J. P. Brown & M. F. Tuite. London:Academic Press.
  13. Heinisch, J. J., Boles, E. & Timpel, C. ( 1996; ). A yeast phosphofructokinase insensitive to the allosteric activator fructose-2,6-bisphosphate. J Biol Chem 271, 15928-15933.[CrossRef]
    [Google Scholar]
  14. Hoffman, C. S. & Winston, F. ( 1987; ). A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57, 267-272.[CrossRef]
    [Google Scholar]
  15. Holyoak, C. D., Stratford, M., McMullin, Z., Cole, M. B., Crimmins, K., Brown, A. J. P. & Coote, P. J. ( 1996; ). Activity of the plasma membrane H+-ATPase and optimal glycolytic flux are required for rapid adaptation and growth of Saccharomyces cerevisiae in the presence of the weak acid preservative, sorbic acid. Appl Environ Microbiol 62, 3158-3164.
    [Google Scholar]
  16. Holyoak, C., Bracey, D., Piper, P. W., Kuchler, K. & Coote, P. ( 1999; ). The Saccharomyces cerevisiae weak-acid-inducible ABC transporter Pdr12 transports fluorescein and preservative anions from the cytosol by an energy-dependent mechanism. J Bacteriol 181, 4644-4652.
    [Google Scholar]
  17. Hunsley, J. R. & Suelter, C. H. ( 1969; ). Yeast pyruvate kinase: kinetic properties. J Biol Chem 244, 4819-4822.
    [Google Scholar]
  18. Krebs, H. A., Wiggins, D., Stubbs, M., Sols, A. & Bedoya, A. ( 1983; ). Studies on the mechanism of the antifungal action of benzoate. Biochem J 214, 657-663.
    [Google Scholar]
  19. Kretschmer, M. & Fraenkel, D. G. ( 1991; ). Yeast 6-phosphofructo-2-kinase: sequence and mutant. Biochemistry 30, 10663-10672.[CrossRef]
    [Google Scholar]
  20. Lambert, R. J. & Stratford, M. ( 1999; ). Weak-acid preservatives: modelling microbial inhibition and response. J Appl Microbiol 86, 157-164.[CrossRef]
    [Google Scholar]
  21. Mellor, J. E., Dobson, M. J., Roberts, N. A., Kingsman, A. J. & Kingsman, S. M. ( 1985; ). Factors affecting heterologous gene expression in Saccharomyces cerevisiae. Gene 33, 215-226.[CrossRef]
    [Google Scholar]
  22. Müller, S., Zimmermann, F. K. & Boles, E. ( 1997; ). Mutant studies of phosphofructo-2-kinases do not reveal an essential role of fructose-2,6-bisphosphate in the regulation of carbon fluxes in yeast cells. Microbiology 143, 3055-3061.[CrossRef]
    [Google Scholar]
  23. Paravicini, G. & Kretschmer, M. ( 1992; ). The yeast FBP26 gene codes for a fructose-2,6-bisphosphatase. Biochemistry 31, 7126-7133.[CrossRef]
    [Google Scholar]
  24. Pearce, A. K., Crimmins, K., Groussac, E., Hewlins, M. J. E., Dickinson, J. R., Francois, J., Booth, I. R. & Brown, A. J. P. ( 2000; ). Pyruvate kinase (Pyk1) levels influence both the rate and direction of carbon flux in yeast under fermentative conditions. Microbiology 147, 391-401.
    [Google Scholar]
  25. Piper, P. W., Mahe, Y., Thompson, S., Pandjaitan, R., Holyoak, C., Egner, R., Muhlbauer, M., Coote, P. & Kuchler, K. ( 1998; ). The Pdr12 ABC transporter is required for the development of weak organic acid resistance in yeast. EMBO J 17, 4257-4265.[CrossRef]
    [Google Scholar]
  26. Ramos, S. M., Balbin, M., Raposo, E. & Pardo, L. A. ( 1989; ). The mechanism of intracellular acidification induced by glucose in Saccharomyces cerevisiae. J Gen Microbiol 135, 2413-2422.
    [Google Scholar]
  27. Reibstein, D., den Hollander, J. A., Pilkis, S. J. & Shulman, R. G. ( 1986; ). Studies on the regulation of yeast phosphofructo-1-kinase: its role in aerobic and anaerobic glycolysis. Biochemistry 25, 219-227.[CrossRef]
    [Google Scholar]
  28. Roe, A. J., McLaggan, D., Davidson, I., O’Byrne, C. & Booth, I. R. ( 1998; ). Perturbation of anion balance during inhibition of growth of Escherichia coli by weak acids. J Bacteriol 180, 767-772.
    [Google Scholar]
  29. Serrano, R. ( 1991; ). Transport across yeast vacuolar and plasma membranes. In The Molecular Biology of the Yeast Saccharomyces: Genome Dynamics, Protein Synthesis and Energetics , pp. 523-585. Edited by J. N. Strathern, E. W. Jones & J. R. Broach. Cold Spring Harbor, NY:Cold Spring Harbor Laboratory.
  30. Thomas, B. J. & Rothstein, R. ( 1989; ). The genetic control of direct-repeat recombination in Saccharomyces: the effect of rad52 and rad1 on mitotic recombination at GAL10, a transcriptional regulation gene. Genetics 123, 725-738.
    [Google Scholar]
  31. Towbin, H., Staehelin, T. & Gordon, J. ( 1979; ). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76, 4350-4354.[CrossRef]
    [Google Scholar]
  32. Van Schaftingen, E., Lederer, B., Bartrons, R. & Hers, H.-G. ( 1982; ). A kinetic study of pyrophosphate: fructose-6-phosphate phosphotransferase from potato tubers. Eur J Biochem 129, 191-195.[CrossRef]
    [Google Scholar]
  33. Viegas, C. A. & Sà-Correia, I. ( 1991; ). Activation of plasma membrane ATPase of Saccharomyces cerevisiae by octanoic acid. J Gen Microbiol 137, 645-651.[CrossRef]
    [Google Scholar]
  34. Wach, A., Brachat, A., Rebischung, C., Steiner, S., Pokorni, K, te Heesen, S. & Philippsen, P. ( 1998; ). PCR-based gene targeting in Saccharomyces cerevisiae. In Yeast Gene Analysis: Methods in Microbiology , pp. 67-81. Edited by A. J. P. Brown & M. F. Tuite. London:Academic Press.
  35. Warth, A. D. ( 1988; ). Effect of benzoic acid on growth yield of yeasts differing in their resistance to preservatives. Appl Environ Microbiol 54, 2091-2095.
    [Google Scholar]
  36. Warth, A. D. ( 1991; ). Effect of benzoic acid on glycolytic metabolite levels and intracellular pH in Saccharomyces cerevisiae. Appl Environ Microbiol 57, 3415-3417.
    [Google Scholar]
  37. Wicksteed, B. L., Collins, I., Dershowitz, A., Stateva, L. I., Green, R. P., Oliver, S. G., Brown, A. J. P. & Newlon, C. S. ( 1994; ). A physical comparison of chromosome III in six strains of Saccharomyces cerevisiae. Yeast 10, 39-57.[CrossRef]
    [Google Scholar]
  38. Yun, S. L., Aust, A. E. & Suelter, C. H. ( 1976; ). A revised preparation of yeast (Saccharomyces cerevisiae) pyruvate kinase. J Biol Chem 251, 124-128.
    [Google Scholar]
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