AbstractThe single frequency ultrasound mode is difficult to achieve an higher enzymolysis efficiency. The cost of protein enzymatic hydrolysis, using free enzyme is higher because the enzyme cannot be used repeatedly. Therefore, the effects of triple-frequency ultrasound (TFU) treatment on the performance, kinetics, and thermodynamics of immobilized Alcalase enzymolysis of corn gluten meal (CGM) were investigated in this research. The results showed that degree of hydrolysis (DH), peptide concentration, ACE inhibitory activity, and relative enzyme activity were increased by 20.6 %, 34.4 %, 24.1 %, and 25.8 %, respectively, by TFU treatment at the optimum conditions compared to the control. Kinetics and thermodynamic analyses revealed that TFU treatment successfully decreased the apparent constant (KM) by 27.0 % and increased the reaction rate constants (k) by 32.1 –200 % at 303.15–343.15 K. The energy of activation (Ea), enthalpy of activation (ΔH), and entropy of activation (ΔS) were reduced by 17.1 %, 15.2 –15.3 %, and 24.1 –31.8 %, respectively. Immobilized enzymolysis assisted by TFU was proved to be an efficient method to increase the enzymolysis efficiency, enzyme activity, and antihypertensive activity of the peptides through performance and mechanism discussion.