Analysis of major phospholipid species and ergosterol in fermenting industrial yeast strains using atmospheric pressure ionization ion-trap mass spectrometry

Knowledge of the individual lipid species that are associated with ethanol tolerance in Saccharomyces cerevisiae is necessary to understand potential mechanisms of how this organism uses these molecules to mitigate the toxic effects of ethanol. Three industrial yeast strains with varying degrees of ethanol tolerance were examined utilizing normal phase high-performance liquid chromatography and atmospheric pressure ionization-ion-trap mass spectrometry methods to quantitatively determine phospholipid and ergosterol levels at numerous fermentation time points. Both high and low Brix fermentations were performed to assess the sugar utilization capabilities of the strains. The results indicated that the strain with the most robust fermentation characteristics had the highest phosphatidylinositol levels and lowest phosphatidylcholine levels. Examination of the phospholipid structural data from tandem MS experiments indicated that the levels of several phospholipid species were unique to the slowest fermenting strain. The relation of ergosterol and other phospholipids to ethanol tolerance is also discussed.