Aim To evaluate the ability of probiotic Saccharomyces cerevisiae RC016 strain to reduce fumonisin B1 (FB1) in vitro and to optimize the culture conditions for the growth of the yeast employing surface response methodology. Methods and Results Using PlackettBurman screening designs (PBSD) and central composite designs (CCD), an optimized culture medium containing (gl1) fermentable sugars provided by sugar cane molasses (CMs), yeast extract (YE) and (NH4)2HPO4 (DAP) was formulated. The S.cerevisiae RC016 strain showed the greatest binding at all assayed FB1 concentration. The CMs, YE, DAP concentrations and incubation time influenced significantly the biomass of S.cerevisiae RC016. Conclusion A combination of CMs 17%; YE 4 center dot 61gl1 and incubation time 60h was optimum for maximum biomass of S.cerevisiae RC016. Significance and Impact of the Study The importance of this work lies in the search for live strains with both probiotic and fumonisin B1 decontamination properties that could be sustainably produced in a medium just containing cheap carbon, nitrogen and phosphorus sources and would be included in a novel product to animal feed.