Use of Tuna Visceral Pepsin in Combination with Trypsin as Digestion Aid: Enhanced Protein Hydrolysis and Bioavailability

Freeze-dried tuna pepsin powder (TPP) was prepared using maltodextrin (10%) and trehalose (5%), while trypsin-loaded beads (TLB) with 5% glycerol were obtained via chitosan/alginate ionotropic gelation. The storage stability of TPP and TLB and their proteolytic activity toward red kidney bean protein (RKB), threadfin bream surimi (TBS) and egg white protein (EWP) in varying simulated in vitro gastrointestinal (GI) tract conditions were studied. The intestinal transepithelial transportation of generated peptides was also carried out through Caco-2 cell monolayers after the cytotoxicity test. Enzyme activity was dropped when TPP and TLB in blister packs were kept for 10 weeks of storage at room (28 °C) and refrigerated (4 °C) temperatures. TPP and TLB at a level of 50% (w/w of proteins) effectively hydrolyzed RKB, TBS and EWP in a simulated in vitro GI tract, as indicated by marked protein degradation and increased degree of hydrolysis. Some peptides generated after GI digestion could transport through Caco-2 cell monolayers. Those peptides had different molecular size distribution and antioxidant activities. The highest antioxidant activity was observed for RKB hydrolysate after passing through the Caco-2 cell monolayer. Therefore, TPP and TLB from skipjack tuna viscera could potentially be used for enzyme supplementation to help digest food proteins. Food-derived bioactive peptides generated after GI digestion could assist in improving human health due to their antioxidant activity.