Metabolic insights into the yeast response to propionic acid based on high resolution 1H NMR spectroscopy
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The experimental model Saccharomyces cerevisiae has been widely used to elucidate the molecular mechanisms behind resistance to weak acids in fungi, an essential knowledge for the development of more suitable preservation strategies. Previous studies, based on transcriptomic and chemogenomic approaches, revealed a number of yeast responses to propionic acid, widely used in the preservation of bakery and fresh dairy products. In the present work we report the metabolic changes occurring during yeast adaptation to, and growth in, the presence of this weak acid (20 mM at pH 4) using high resolution 1H NMR spectroscopy coupled with multivariate statistical analysis. The metabolic profiles highlighted the separation of propionic acid-induced lag-phase in two parts. The initial period of incubation under acid stress (up to 3 h following the inoculation of an unadapted yeast population) was characterized by a decrease of cell viability and of the average intracellular pH (pHi) values. The final part of this incubation period (from 4 to 6 h of incubation) was characterized by the start of cell division in the presence of the acid, an increase of the average pHi and a metabolic profile close to the profile exhibited by cells in the exponential phase of growth in propionic acid supplemented medium. An association between the average pHi values and the levels of glutamate and propionate during growth latency was identified. Changes in the cell content in other amino acids, ATP, NAD+, glycerol and trehalose were also registered during yeast incubation with propionic acid. These alterations are discussed in the context of the global response to this weak acid.
KeywordsSaccharomyces cerevisiae Response to propionic stress Weak acids 1H NMR-based metabolomics Multivariate data analysis
This research was supported by FEDER, Fundação para a Ciência e a Tecnologia (FCT) (PTDC/AGR-ALI/102608/2008 grant and a PhD fellowship grant to ABL/SFRH/BD/23437/2005). The Portuguese National NMR Network is acknowledged for providing us the NMR facility.
- Fernandes, A. R., Durão, P. J., Santos, P. M., & Sá-Correia, I. (2003). Activation and significance of vacuolar H+-ATPase in Saccharomyces cerevisiae adaptation and resistance to the herbicide 2,4-dichlorophenoxyacetic acid. Biochemical and Biophysical Research Communications, 312, 1317–1324.PubMedCrossRefGoogle Scholar
- Heinisch, J. (1986). Isolation and characterization of the two structural genes coding for phosphofructokinase in yeast. Molecular Genetics and Genomics, 202, 75–82.Google Scholar
- Holyoak, C. D., Stratford, M., McMullin, Z., Cole, M. B., Crimmins, K., Brown, A. J. P., et al. (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. Applied and Environmental Microbiology, 62, 3158–3164.PubMedGoogle Scholar
- Larsson, C., Nilsson, A., Blomberg, A., & Gustafsson, L. (1997). Glycolytic flux is conditionally correlated with ATP concentration in Saccharomyces cerevisiae: a chemostat study under carbon- or nitrogen-limiting. Conditions Journal of Bacteriology, 179, 7243–7250.Google Scholar
- Lundberg, P., Vogel, T., Malusek, A., Lundquist, P. O., Cohen, L., & Dahlqvist, O. (2005). MDL—the magnetic resonance metabolomics database (mdl.imv.liu.se). Basel, Switzerland: ESMRMB.Google Scholar
- Otto, A., Przybylski, F., Nissler, K., Schellenberger, W., & Hofmann, E. (1986). Kinetic effects of fructose-1, 6-bisphosphate on yeast phosphofructokinase. Biomedicine Biochimica Acta, 45, 865–875.Google Scholar
- Pears, M. R., Codlin, S., Haines, R. L., White, I. J., Mortishire-Smith, R. J., Mole, S. E., et al. (2010). Deletion of btn1, an orthologue of CLN3, increases glycolysis and perturbs amino acid metabolism in the fission yeast model of Batten disease. Molecular BioSystems, 6, 1093–1102.PubMedCrossRefGoogle Scholar
- Roe, A. J., Mclaggan, D., Davidson, I., O’Byrne, C., & Booth, I. R. (1995). Perturbation of anion balance during inhibition of growth of Escherichia coli by weak acids. Journal of Bacteriology, 180, 767–772.Google Scholar
- Sá-Correia, I., Salgueiro, S. P., Viegas, C. A., & Novais, J. M. (1989). Leakage induced by ethanol, octanoic and decanoic acids in Saccharomyces cerevisiae. Yeast, Special Issue, 5, S123–S127.Google Scholar
- Viegas, A. C., Almeida, P. F., Cavaco, M., & Sá-Correia, I. (1998). The H+-ATPase in the plasma membrane of Saccharomyces cerevisiae is activated during growth latency in octanoic acid-supplemented medium accompanying the decrease in intracellular pH and cell viability. Applied and Environmental Microbiology, 64, 779–783.PubMedGoogle Scholar