Although vitamins are prime actors in yeast metabolism, the nature and the extent of their requirement in Saccharomyces cerevisiae in winemaking remains little understood. To fill this gap, the evolution of 8 water-soluble vitamins and their diverse vitamers during its alcoholic fermentation in a synthetic must medium was monitored, providing the first evidence of the consumption of vitamers by five commercial S. cerevisiae strains, and highlighting the existence of preferential vitameric sources for its nutrition. The vitamins required by the yeast, B1, B5, and B8, were then identified, and the nature of their requirement characterized, strongly asserting the required trait of B1 for fermentation, B8 for growth, and B5 for both processes. The extent of the requirement for B5, that with the most impact of the three vitamins, was then quantified in three S. cerevisiae strains, resulting in the conclusion that 750 μg.L-1 should prove sufficient to cover the yeast's requirements. This investigation offers the first insight into S. cerevisiae vitaminic requirements for winemaking.