In view of utilizing fish viscera wastes as source of both, protein and proteases, hydrolysates from smooth hound viscera were prepared using endogenous enzymes, commercial proteases and a combination of these two preparations. Their antioxidant, ACE-inhibitory and antimicrobial activities were studied. The co-digestion with endogenous enzymes in combination with commercial proteases was found to enhance protein hydrolysis. The resulted smooth hound viscera hydrolysates (SHVHs) were mainly constituted by Gly, Glu and Gln, while Tau was found the major free amino acid. By contrast to the undigested proteins containing high amounts of IMP and Xanthine, the UMP, Uridine, GMP and Guanosine were the major nucleotides derivates detected in the SHVHs. Furthermore, SHVHs showed distinct molecular mass distribution and RP-HPLC profiles proving their molecular mass and hydrophilic/hydrophobic peptide heterogeneity. All the SHVHs exhibited important antioxidant activities in terms of radical-scavenging activity, reducing power, metal chelating activity, β-carotene protection, lipid peroxidation inhibition and DNA breakage assay. Additionally, they possessed considerable antibacterial effect against several strains. Further, all hydrolysates showed varying degrees of ACE inhibitory activities and the highest one was achieved by purafect hydrolysate (IC 50 = 75 µg/ml). The overall data suggested that the SHVHs could be used as potential source of natural antioxidant, antimicrobial and anti-ACE peptides to formulate functional foods.