To utilize low-value ﬁsh, Spanish mackerel protein was hydrolyzed using alcalase for production of antioxidant peptide. The antioxidant activities of antioxidant peptide from Spanish mackerel protein were evaluated. Results: The radical-scavenging properties on hydroxyl and superoxide of protein hydrolysates were 53.11% and 23.1%. And their reducing power was increased with the peptide content of the hydrolysates. The hydrolysates signiﬁcantly inhibited lipid peroxidation in linoleic acid emulsion system, and effectively inhibit autoxidation of linoleic acid; and the inhibition ability increased with the increasing of peptide content in hydrolysates. The hydrolysates inhibited the activity of LOX to a certain extent. Conclusion: The hydrolysates from Spanish mackerel protein exhibited high antioxidative activity.