This work allowed the evaluation of the gastrointestinal resistance of five yeasts (Saccharomyces and non-Saccharomyces) in order to assess some biotechnological characteristics linked to the potential probiotics, using a dynamic gastrointestinal simulator (simgi®). The best results obtained were for strains Saccharomyces cerevisiae 3 and Hanseniaspora osmophila 1056. Having optimised the method, the yeasts were subsequently lyophilised, and the one that showed the least loss of viability, S. cerevisiae 3, was used in a freeze-dried form to obtain a new functional food. On the other hand, some characteristics of the product were compared with those of probiotic supplements and other commercial probiotic foods. The obtained functional product showed better parameters than the rest of the samples containing yeasts which, together with the great acceptance shown after the consumer tests, means that it can be presented as a possible commercial functional product.