An innovative perspective on fermented foods: isolation, purification, biochemical properties, and evaluation of the flavor formation potential of meat protein hydrolysis by Saccharomyces cerevisiae L3 proteases.

Abstract

This study evaluated the extracellular protease characteristics, protein hydrolysis activity, and flavor formation of Saccharomyces cerevisiae L3. The protease was purified using ammonium sulfate precipitation, size exclusion chromatography, and molecular sieve chromatography, yielding a relative molecular weight of 48.0 kDa. The protease exhibited maximum activity at pH 8 and 50 °C. The presence of Mg²⁺ and Ca²⁺ significantly enhanced the activity of protease (P < 0.05). The activity of protease was significantly inhibited by ethylene diamine tetraacetic acid (P < 0.05). The K_m and V_max of the protease were 17.84 mg/mL and 19.61 U/mL, respectively. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis revealed the protease's ability to hydrolyze myofibrillar proteins (MPs). Moreover, as protease hydrolysis progresses, the solubility of MPs, the levels of free amino acids, and volatile compounds all increase, thereby enhancing the flavor. The results demonstrated the potential application of S. cerevisiae protease in fermented meat products.