The Effect of Yeast Inoculation Methods on the Metabolite Composition of Sauvignon Blanc Wines

Evidence from the literature suggests that different inoculation strategies using either active dry yeast (ADY) or freshly prepared yeast cultures affect wine yeast performance, thus altering biomass and many primary and secondary metabolites produced during fermentation. Here, we investigated how different inoculation methods changed the fermentation behaviour and metabolism of a commercial wine yeast. Using a commercial Sauvignon blanc (SB) grape juice, fermentation was carried out with two different inoculum preparation protocols using <i>Saccharomyces cerevisiae</i> X5: rehydration of commercial ADY and preparation of pre-inoculum in a rich laboratory medium. We also determined the effect of different numbers of yeast cells inoculation (varying from 1 × 10⁶ to 1 × 10¹²) and successive inoculation on fermentation and end-product formation. The yeast inoculation method and number of cells significantly affected the fermentation time. Principal component analysis (PCA) using 60 wine metabolites showed a separation pattern between wines produced from the two inoculation methods. Inoculation methods influenced the production of amino acids and different aroma compounds, including ethyl and acetate esters. Varietal thiols, 3-mercaptohexanol (3MH), and 4-methyl-4-mercaptopentan-2-one (4MMP) in the wines were affected by the inoculation methods and numbers of inoculated cells, while little impact was observed on 3-mercaptohexyl acetate (3MHA) production. Pathway analysis using these quantified metabolites allowed us to identify the most significant pathways, most of which were related to central carbon metabolism, particularly metabolic pathways involving nitrogen and sulphur metabolism. Altogether, these results suggest that inoculation method and number of inoculated cells should be considered in the production of different wine styles.