In red winemaking, polyphenols from grape berry pericarp and seed are extracted during fermentation and their interactions with yeast have been widely demonstrated. However, information concerning the impact of extracted polyphenols on yeast metabolism during fermentation is missing. The aim of this study was to further explore interactions between yeasts and polyphenols and to identify their effects on yeast metabolism and fermentation kinetics. This impact was studied in synthetic musts for four commercial Saccharomyces cerevisiae wine strains, using polyphenols purified from a thermovinification must, in both stressed (phytosterol deficient medium) and non-stressed conditions. Interactions between grape polyphenols and yeast cells were substantiated from the early stage of fermentation by means of epifluorescence and confocal microscopy. If these interactions were limited to yeast cell walls in non-stressed conditions, the passage of polyphenols through yeast envelope and their accumulation in the intracellular space of living cells was shown in phytosterol-deficient medium. Whatever the conditions used (stressed and non-stressed conditions) and for all strains, the presence of polyphenols led to a significant decrease of cell growth (50%), CO2 production rate (60 to 80%) and nitrogen consumption (3 to 4 times less), resulting in increased fermentation lengths. The perturbation of yeast growth and metabolism due to polyphenol compounds was likely mostly linked to their interactions with the yeast plasma membrane. From the mid-stationary phase to the end of the fermentation, an adaptive response was exhibited by yeast, resulting in lower mortality. This work evidenced a strong impact of polyphenols on yeast fermentative capacity and highlighted the importance of a better knowledge of the mechanisms involved to improve the management of fermentations in the context of red winemaking.