Response Surface Modeling and Optimization of Enzymolysis Parameters for the In Vitro Antidiabetic Activities of Peanut Protein Hydrolysates Prepared Using Two Proteases

Abstract

Optimization of the enzymolysis process for preparing peanut protein hydrolysates using alcalase and trypsin was performed by employing the central composite design (CCD) of response surface methodology (RSM). The independent variables were solid-to-liquid ratio (S/L), enzyme-to-substrate ratio (E/S), pH, and reaction temperature, while the response variables were degree of hydrolysate (DH), α-amylase, and α-glucosidase inhibitory activity. The highest DH (22.84% and 14.63%), α-amylase inhibition (56.78% and 40.80%), and α-glucosidase inhibition (86.37% and 86.51%) were obtained under optimal conditions, which were S/L of 1:26.22 and 1:30 w/v, E/S of 6% and 5.67%, pH of 8.41 and 8.56, and temperature of 56.18 °C and 58.75 °C at 3 h using alcalase (AH) and trypsin (TH), respectively. Molecular weight distributions of peanut protein hydrolysates were characterized by SDS-PAGE, which were mostly ˂10 kDa for both hydrolysates. Lyophilized AH and TH had IC50 values of 6.77 and 5.86 mg/mL for α-amylase inhibitory activity, and 6.28 and 5.64 mg/mL for α-glucosidase inhibitory activity. The IC50 of AH and TH against DPPH radical was achieved at 4.10 and 3.20 mg/mL and against ABTS radical at 2.71 and 2.32 mg/mL, respectively. The obtained hydrolysates with antidiabetic activity could be utilized as natural alternatives to synthetic antidiabetics, particularly in food and pharmaceutical products.