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Methodologies: S. boulardii is a probiotic yeast related to Saccharomyces cerevisiae (S. cerevisiae), but with distinct genetic, taxonomic and metabolic properties. S. cerevisiae has been extensively used in biotechnological applications. Currently, many strains are available and multiple genetic tools have been developed, which allow the expression of several exogenous proteins of interest with applications in the fields of medicine, biofuels, food industry and scientific research, among others. Although S. boulardii has been widely studied due to its probiotic properties against several gastrointestinal tract disorders, very few studies addressed the use of this yeast as a vector for expression of foreign genes of interest with biotechnological applications. Here we applied to the probiotic yeast a simple plasmid transformation technique based on the lithium acetate protocol used for S. cerevisiae with minor modifications. We also tried different protocols for screening based on the protoplast formation. We further screened by PCR the genome of S. boulardii for the presence of promoters and other molecules of interest (PGK1, PYK1, ENO1, ADH1, REP1, REP2, CAN1, TAF10, AGA1 and δ transposons).

Saccharomyces boulardii as an alternative expression system for molecules with biotherapeutical applications