In this study, proteins were extracted from tomato seeds, the main by-product of tomato processing. The incubation for 138.62 min coupled with 3% alcalase was observed to be optimum to produce a tomato seed protein hydrolysate (TSPH) with the highest antioxidant properties. Under these conditions, predicted TSPH activities were 62.99% scavenging of DPPH radicals and 54.81% reduction of phosphomolybdate. Separation of TSPH by ultrafiltration provided three fractions (UF1-UF3) of which, UF3 (< 3 kDa) showed the strongest activity (73.15% DPPH scavenging and 60.1% phosphomolybdate reduction). UF3 was further separated by RP-HPLC into sub-fractions F1-F6. Biological testing found that F2 and F4 were the most active in scavenging DPPH radicals (60.36 and 21.23%) and reducing phosphomolybdate (57.3 and 48.0%). LC-ESI-MS/MS analysis showed that the higher activity of F2 might be explained by the presence of more peptides that contained tyrosine and histidine, known to enhance antioxidant activity through hydrogen or electron transfer. In the simulated gastrointestinal digestion test, peptides in F2 were more resistant compared to those in F4. These findings indicate that peptide fraction F2 might be more useful in the formulation of functional foods because of its greater antioxidant activity and resistance to digestion.
Keywords: Antioxidant peptides; Gastrointestinal digestion; LC–ESI–MS/MS; Ultrafiltration.