Identification of antioxidative oligopeptides derived from autolysis hydrolysates of sea cucumber (Stichopus japonicus) guts

摘要

Oligopeptides were prepared from the guts of sea cucumber Stichopus japonicus by autolysis method. Optimum autolysis conditions for preparing oligopeptides from the guts of sea cucumber were determined by response surface methodology using a central composite rotatable design. The effects of two independent variables, namely temperature and pH, on the response of trichloroacetic acid-soluble oligopeptides (mg/g on dry basis) were investigated. Regression analysis indicated that more than 95 % of the variation could be explained by the fitted models. Temperature at 48.30 °C and pH at 4.43 were found to be the optimal conditions to obtain oligopeptides from the guts of sea cucumber. The autolysis hydrolysates prepared at the optimized conditions were further fractionated into four major fractions (I–IV) by size exclusion chromatography on a Sephadex G-15 column. Fraction IV, which exhibited the highest DPPH radical scavenging capacity, Fe2+-chelating ability and protective effect against hydroxyl radical-induced DNA damage, was then analyzed by ESI–MS for molecular mass determination and ESI–MS/MS for the characterization of peptides. Two tetrapeptides (Val-Thr-Pro-Tyr and Val-Leu-Leu-Tyr) and a hexapeptide (Val-Gly-Thr-Val-Glu-Met) were found to exhibit protective effects against hydroxyl radical-induced DNA damage. These results suggest that antioxidant oligopeptides derived from the guts of sea cucumber by autolysis method could be utilized for functional foods.