This study was conducted to obtain the gelatin fraction with a high anti oxidative activity from Alaska pollock surimi by-products using a two-step enzymatic hydrolysis and ultrafiltration. Among gelatin hydrolysates from refiner discharge of Alaska Pollock surimi, the highest antioxidative activity (81.5%) resulted from gelatin hydrolysate sequentially treated with Pronase E and Flavourzyme each for 2 hr. However, no difference was seen in the anti oxidative activity of the second hydrolysate (Pronase E-/Flavourzyme-treated hydrolysate) when compared to the permeate fractionated through a 10-kDa membrane. The results suggest that the Pronase E-/Flavourzyme-treated hydrolysate from refiner discharge gelatin of Alaska pollock surimi can be used as a supplementary raw material for improving health functionality.