Bioactive peptides derived from food have been increasingly popular due to their therapeutic properties. Of particular importance are peptides with a multidirectional activity that can be used in the treatment and prevention of diet-related diseases. This paper attempts to utilize a by-product of phospholipid extraction from egg yolk as a source of peptides with a broad spectrum of biological activity. In addition, in this research we used a non-commercial enzyme obtained from Asian pumpkin, which has not been sufficiently researched in terms of its ability to release biopeptides from food proteins. In the present study the biological properties of peptides, derived from egg-yolk protein by-products (YP) remaining after phospholipid extraction, and their four synthetic analogs were investigated with regard to their antioxidant (radical scavenging capacity, Fe 2+ chelating effect, reducing power (FRAP)) and antidiabetic (a-glucosidase and DPP-IV inhibitory activities) properties. One of them, with the sequence LAPSLPGKPKPD, exhibited the highest antioxidant activity (free radical scavenging activity (6.03 mM Trolox eq per mg protein); FRAP (296.07 mg Fe 2+ per mg protein)). This peptide also revealed the strongest DPP-IV (361.5 mmol L À1 ) and a-glucosidase (1065.6 mmol L À1 ) inhibitory activities, a novel multifunctional effect for peptides from an egg-yolk hydrolysate.