Introduction: Saccharomyces cerevisiae has been widely used for fermenting food and beverages for over thousands years. Its metabolism together with the substrate composition play an important role in determining the characteristics of the final fermented products. We previously showed that the polyunsaturated fatty acid, linoleic acid, which is present in the grape juice at trace levels, significantly affected the development of aroma compounds of the wines. However, the effect of linoleic acid on the overall cell metabolism of S. cerevisiae is still not clear. Therefore, we aimed to unlock the metabolic response of S. cerevisiae to linoleic acid using metabolomics and isotope labelling experiments.
Methods: We cultured the cells on a minimal mineral medium supplementing them with linoleic acid isomers and 13C-linoleic acid. Both intracellular and extracellular metabolite profiles were determined using gas chromatography coupled to mass spectrometry (GC-MS) to investigate which S. cerevisiae pathways were affected by linoleic acid supplementation.
Results: The utilisation of linoleic acid by S. cerevisiae had a significant impact on the primary carbon metabolism increasing the glucose consumption and the ethanol production under anaerobic condition. The energetic state of the cell was, therefore, affected and the glycolytic pathway, the TCA cycle and the amino acid production were up-regulated. We also observed that linoleic acid was transported into the cell and converted into other fatty acids affecting their profile even under anaerobic condition.
Conclusion: Our data clearly shows that linoleic acid supplementation in growth medium increased glucose consumption and ethanol production by S. cerevisiae under anaerobic condition. We also suggest that S. cerevisiae might be able to perform an alternative anaerobic pathway to β-oxidation, which has not been reported yet.
Keywords: Conjugated linoleic acid; Ethanol; Extracellular metabolites; Gas-chromatography and mass-spectrometry; Intracellular metabolites; Isotope labelling; Pathway analysis.