Hybrids of the homothallic diploid Saccharomyces mikatae and heterothallic haploid Saccharomyces cerevisiae isolated from local fruits in Hokkaido and a baking strain, respectively, were constructed by the spore to cell mating method. Interspecific hybridization of the selected strain was confirmed by polymerase chain reaction using species-specific primers, fermentation profiles, and spore formation. The leavening ability of the dough and enzyme activities of the hybrid exhibited intermediate traits of their parental strains. A baking test by the sponge-dough method with a standard formulation of white bread showed that breads produced using the hybrid differed in aroma and taste from those made using a conventional baking strain. Thus, the hybrid may be used to produce baked goods with diverse quality, which may be more acceptable to specific consumers.