Loach (Misgurnus anguillicaudatus) proteins were hydrolysed by papain and Protamex, the antioxidant activity of loach protein hydrolysates (LPH) was investigated. The results demonstrated that extensive hydrolysis by papain and Protamex led to the browning of the hydrolysates. When the degree of hydrolysis (DH) was 23%, hydrolysates prepared by papain (HA) exhibited the strongest antioxidant activity. The maximum values of the hydroxyl, 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-etllylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging activities and the reducing power were 56.1%, 95.5%, 2.80 mM and 1.46, respectively. The hydrolysates prepared by Protamex (HB) showed the strongest hydroxyl radical scavenging activity (55.0%) at DH 28%, DPPH radical scavenging activity (92.2%) and ABTS radical scavenging activity (2.81 mM) at DH 23%, and the reducing power (1.17) at DH 33%. At the same DH value, there were significant (p<0.05) differences between HA and HB. Several antioxidant amino acid residues, especially Trp and His, contributed significantly to the antioxidant activity of the hydrolysates. An increase of peptides with molecular weight below 500 Da was observed as the DH increased for all LPH. The above results indicated that DH and protease greatly influenced the molecular weight and amino acid residue composition of LPH, and further influenced the antioxidant activity. Industrial relevance: Loach has long been employed as a traditional Chinese medicine for the treatment of many kinds of diseases. From our previous work, leach was determined to be a good source of protein (accounts for approximately 17% (w/w) of the body weight). In this work, loach proteins were hydrolyzed by papain and Protamex to specific extent. The effect of DH on the antioxidant activities of hydrolysates was investigated. The results indicated that leach protein hydrolysates were potent antioxidants which were significantly affected by DH. This research is helpful for extensive development of loach product. Crown Copyright (C) 2008 Published by Elsevier Ltd. All rights reserved.