The probiotic potential of Leuconostoc mesenteroides subsp. mesenteroides SJRP55, isolated from water buffalo mozzarella cheese was evaluated. The microorganism presented resistance to stressful conditions that simulated the gastrointestinal tract, and to the best of our knowledge, Leuconostoc mesenteroides subsp. mesenteroides SJRP55 was the first of this species with the ability to deconjugate bile salts. Tolerance to NaCl was temperature dependent, as well the results obtained by aggregation capacity. The strain presented good adhesion properties, β-galactosidase activity, viability in fermented milk during storage, inactive against Streptococcus thermophilus and sensitive to most of the tested antibiotics. Some analgesic medications inhibited the growth of the strain. Leuconostoc mesenteroides subsp. mesenteroides SJRP55 exhibited in vitro probiotic potential, and it can be better characterized through future in vivo tests. This bacterium presents higher functional properties compared to other studied strains, and therefore, it is a potential candidate for the application as a probiotic strain, which could be used by industries in the manufacture of functional milk-based products.