In this study, we investigated the bioactive substances of the Alcalase hydrolysate obtained from fishery processing by-products in Jeju by measuring bioactivities including radical scavenging acitivty, cytoprotective activity against 2,2-azobis-(2-amidino-propane) dihydrochloride(AAPH), and ACE inhibitory activity. This study is important because of utilization of unused fishery processing by-products in Jeju. The Alcalase hydrolysate was prepared through the hot water extraction and enzymatic hydrolysis, and then further separation of the Alcalase hydrolysate was performed by ultrafiltration using 10 kDa molecular weight cut-off membrane. The Alcalase hydrolysate showed the relatively higher DPPH and peroxyl radical scavenging activity (IC<SUB>50</SUB> value; 1.30 mg/ml and 0.888 mg/ml, respectively). Also, the Alcalase hydrolysate showed the ACE inhibitory activity with 1.87 mg/ml of IC<SUB>50</SUB> value. These biological activities are increased over 1.2 or 2.5 times through the ultrafiltration of the Alcalase hydrolysate. Therefore, the Alcalase hydrolysate obtained from fishery processing by-products in Jeju and the different molecular weight fractions should be given consideration for food and cosmetics ingredient. Furthermore, this research on the utility of fishery processing by-products might be a useful tool into the industry.