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Protein hydrolysis of underutilized marine organisms is a method to improve added value. In the present article, anchovy protein was autolyzed at pH 8.5 and 50°C. After autolysis processing for 24 h, total nitrogen recovery reached 92.1%. The protein hydrolysates had antioxidant properties on reducing power, 1, 1-diphenyl-2-picryhydrazyl radical, and hydroxyl radical scavenging activity. More than 79% of the peptides in the hydrolysates had a molecular weight of lower than 3 kDa. Protein hydrolysates had balanced amino acid compositions, and total content of essential amino acids had no significant difference after hydrolysis. Protein hydrolysates from anchovy autolysis have potential in nutritional and functional foods.