Saccharomyces cerevisiae var. boulardii is the only yeast indicated as a probiotic. In recent decades, sports drinks have become very popular due to their peculiarities of providing health benefits; however, the supplementation of probiotics in beverages or foods requires to address at least two challenges, that is the production of biomass and the viability of probiotics in the product, along with the possible effect on the sensory scores. Therefore, the main topic of this paper, was to study the effects of adding two commercial strains of S. boulardii, labeled as SB1 and SB2, to a beverage intended for athletes by focusing on their viability and consumers’ preference, while as a preliminary step for biomass production the effects of pH, glucose concentration and temperature were studied through the methodology of Central Composite Design. Concerning biomass production, the combination of both response surface methodology and desirability approach highlights the existence of non-linear trends for the effects of temperature and glucose on the growth kinetic, with optimal values depending on the time, that is 30-35°C and 10-15 g/L of glucose after 24 h and 25°C and 15-20 g/L of glucose after 48 h, while pH was less or not significant. Both probiotic yeasts survived in the beverage, although consumers showed a preference for the strain SB2, thus suggesting that the interaction probiotic microorganism/food should be carefully assessed and evaluated in each case.