Inoculation-density-dependent responses and pathway shifts in Saccharomyces cerevisiae.

The cell-density-dependent responses of Saccharomyces cerevisiae to inoculation sizes were explored by a proteomic approach. According to their gene ontology, 100 protein spots with differential expression, corresponding to 67 proteins, were identified and classed into 17 different functional groups. Upregulation of eight heat shock, oxidative response and amino acid biosynthesis-related proteins (e.g. Hsp78p, Ssa1p, Hsp60p, Ctt1p, Sod1p, Ahp1p, Met6p and Met17p), which may jointly maintain the cell redox homeostasis, was dependant on inoculation density. Significant increases in the levels of five proteins involved in glycolysis and alcohol biosynthesis pathways (e.g. Glk1p, Fba1p, Eno1p, Pdc1p and Adh1p) might play critical roles in improving ethanol productivity of the fermentation process and shortening the fermentation time when inoculation sizes were increased. Cell-density-dependent glycolytic variations of proteins involved in trehalose, glycerol biosynthesis and pentose phosphate pathway revealed shifts among metabolic pathways during fermentation with different inoculation sizes. Upregulation of three signal transduction proteins (Bmh1p, Bmh2p and Fpr1p) indicated that adequate cell-cell contacts improved cellular communication at high inoculation sizes. These findings provide insights into yeast responses to inoculation size and optimizing the direct inoculation of active dry yeast fermentation, so as to improve the ethanol production.