Prato cheese, a traditional semi-hard Brazilian cheese is a dairy food largely manufactured for dairy companies. However, the high sodium level Prato cheese formulation constitutes a negative aspect for consumers with present cardiovascular disease. This study aimed to evaluate the effect of sodium reduction and flavor enhancers on the availability of calcium, magnesium, zinc, phosphorus, and potassium of probiotic Prato cheese formulations (L. casei 01, 6 log cfu/mL). The cheese was manufactured considering 5 different formulations: 100% NaCl, NaCl/KCl 50/50%, and NaCl/KCl 50/50% wt/wt using the following flavor enhancers (oregano extract, arginine, and yeast extract, respectively). Mineral content and availability were evaluated at 3 different periods: 1, 30 and 60 d (immediately after the processing, ripening and refrigerated storage). Sodium reduction and addition of flavor enhancers influenced the mineral content and availability (P > 0.05), being the performance dependent of the ingredients added. Oregano extract addition increased the calcium, magnesium, zinc, phosphorus and potassium content (195.63, 146.62, 7.61, 528.36, 367.11 mg/100g, respectively) and their availability (19.78, 65.85, 19.25, 9.22, 8,55%, respectively) after the ripening (60 d), being followed by yeast extract and arginine addition. On the converse, conventional Prato cheese (normal sodium level and without flavor enhancer had lower calcium, magnesium, zinc, phosphorus and potassium content (99.59, 112.20, 3.18, 214.38, 84.72 mg/100g, respectively) and lower availability values (10.63, 52.65, 7.4, 4.32, 8.01%, respectively). Overall, the reformulation of Prato cheese by decreasing sodium content and addition of flavor enhancers increased the mineral content and availability and should take in account for the Brazilian cheese industry.