This study investigated the biotechnological potential of eleven Saccharomyces cerevisiae strains (1P–11P) isolated from artisanal and commercial sourdoughs originating from the Republic of Moldova, Romania, and Ukraine. The strains were cultivated under laboratory conditions on YPD medium, and several physiological, biochemical, and technological parameters were evaluated, including biomass production, protein and carbohydrate content, antioxidant activity, enzymatic activity, and dough leavening performance. The obtained results demonstrated significant differences among the strains (p ≤ 0.05). Biomass production ranged from 2.73 to 6.53 g·L-1, with the highest value recorded for strain 6P. Protein content varied between 41.02% and 57.92% dry weight, with strains 2P and 4P showing the highest levels. The strongest total antioxidant activity was observed in strains 1P and 2P. Superoxide dismutase activity reached its maximum in strains 3P and 4P, while catalase activity was highest in strain 9P, isolated from commercial sourdough. Regarding technological performance, strains 9P and 10P exhibited superior dough leavening ability, characterized by rapid fermentation and efficient CO₂ retention within 90–120 minutes. Overall, artisanal sourdough isolates demonstrated better performance than commercial strains in most evaluated parameters, highlighting their potential for sustainable applications in baking and microbial biotechnology.