<p>The present research work aims to purify and characterize phytase derived from <i>Aspergillus niger</i> NT7, a strain previously identified in our laboratory and to corroborate its potential to hydrolyze anti-nutrients in plant-based food products. Crude phytase, produced via solid-state fermentation using wheat bran, was purified through sequential ammonium sulphate precipitation (85% saturation), dialysis and DEAE-Sepharose anion exchange chromatography, resulting in an 11.48-fold purification and a specific activity of 34.93 U/mg. The purified phytase, having a molecular weight of ~ 70 kDa, exhibited optimal activity at pH 5.0 and 50 °C. The observed kinetic parameters, K_M and V_max values for sodium phytate, were 1.98 mM and 99 U/mg, respectively. The purified phytase demonstrated industrially desired characters, including thermotolerance at 50 °C (t_1/2 288.75 min), 60 °C (t_1/2 106.61 min) and 70 °C (t_1/2 52.90 min) with strong resistance to proteolytic degradation. Phytase efficiently hydrolyzed insoluble protein-phytate and metal-phytate complexes, optimally at 50 °C. This study is the first report to elucidate the application of phytase in dephytinization under simulated digestive conditions, enhancing the nutritional and mineral contents of sorghum flour. Phytase supplementation led to a ~ 90% reduction in phytic acid and significantly enhanced the release of inorganic phosphate (Pi), reducing sugars, proteins and free amino acids from the sorghum flour during simulated digestive conditions of both gastric and intestinal phases. ICP-MS analysis further confirmed enhanced mineral bioaccessibility, with notable increases in cationic metal ions such as Zn²⁺, Fe²⁺ and Mn²⁺. The exclusive characteristics of <i>A. niger</i> NT7 phytase, including acidic resilience, thermotolerance and robust proteolytic resistance, validate its commercial potential as a functional food supplement.</p> Graphical abstract <p></p>