Peptides of marine animals as a potential source of natural antioxidants

Peptides extracted from tissues of 11 shellfish species ( Corbicula japonica, Mercenaria stimpsoni, Anadara broughtoni, Patinopecten yessoesis, Crenomytilus grayanus, Spisula sachalinensis, Maktra chinensis, Serripes groenlandicus, Glycymeris yessoensis, Callista brevisiphonata, Dosinia japonica ) are tested for their antioxidant properties in different in vitro conditions. Frozen muscles and viscera of the shellfish were minced and subjected to water extraction of proteins and to hydrolysis with Protamex 1.5 MG. The peptides antioxidant activity was evaluated by assessing of 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity on free radicals generated in oxidative systems. In the experiment, the high perfomance liquid chromatography (HPLC) system (Agilent Technologies 1260) included the frame TSKgel G 3000PWXL, the flow rate was 0.1 mL/min (0.1 N NaCI-20 mM Tris-HCI, pH 8.0), the detection was made under 280 nm. Molecular weight of the proteins was determined by comparison of their retention time with pure protein standards. The enzyme activity was high in conditions of pH in the range of 5.5-8.0, with the maximum under pH 7.0. Among 11 shellfish species, the highest antioxidant activity in water extract was observed for C. japonica , M. stimpsoni , C. brevisiphonata , G. yessoensis , and C. grayanus : 86.3, 68.7, 72.3, 90.2, and 67.5 mg of ascorbic acid/g, respectively. The DPPH radical scavenging activity increased with increasing of the hydrolysis degree. The 1-step hydrolysis with Protamex enhanced the DPPH scavenging activity for all samples, with the highest value for the protein hydrolysate of M. stimpsoni tissues (215 mg of ascorbic acid/g). Size of generated peptides is important for the antioxidant activity therefore molecular weight distribution of the peptides during the proteolysis was investigated using HPLC. Portion of the low molecular weight peptides (≤ 1 kDa) in the protein hydrolysate was increased in 4-14 % as compared with the starting protein. The antioxidant activity correlated positively with the number of low molecular weight peptides in protein hydrolysates.