Continuous Production of Casein-based Bioactive Peptide using Dual Enzymatic Proteolysis

Abstract

Sodium Caseinate is a derivative form of Casein Protein that is commonly found in milk proteins and has the potential as a parent protein for producing bioactive peptides. The functionality of bioactive peptides can be increased by dual enzyme hydrolysis with alcalase and neutrase enzymes. Several parameters were considered in conducting continuous dual enzyme hydrolysis, namely; enzyme-to substrate concentration ratio (E/S), pH value, and residence time. This research aimed to produce bioactive peptides from sodium caseinate continuously using an enzymatic membrane reactor and study the optimum parameters that can enhance the functionalities of the hydrolysates. Permeate with an E/S of 7.5%, pH value of 7.0, and residence time = 9 h had the highest values for the antioxidant capacity of 19.65 ± 1.81 mgAEAC/mL for DPPH Method, 22.47 ± 6.50 mgAEAC/mL for FRAP method, and 91.39 ± 3.13% inhibition for ABTS method. The value of IC50 antioxidant with the DPPH method was 0,18 ± 0,03 g/mL obtained by filtering permeate with a PES (Polyethersulfone) membrane MWCO of 4 kDa. Keywords: Alcalase, bioactive peptides, casein, enzymatic membrane reactor, neutrase