Novel antioxidant peptides from protein hydrolysates of scallop (Argopecten irradians) mantle using enzymatic and microbial methods: Preparation, purification, identification and characterization

The aim of this study was to isolate, purify, identify, and characterize novel antioxidant peptides from protein hydrolysates of scallop (Argopecten irradians) mantle. Antioxidant peptides were obtained following protein hydrolysis, using a neutral protease, and microbial fermentation which was carried out by Bacillus licheniformis CICC 20033 under optimized conditions. Antioxidant peptides from the scallop mantle hydrolysates were then purified through fractionation using ultrafiltration membranes, and the fraction (<3 kDa) showed the highest oxygen radical absorbance capacity (ORAC) with the value of 0.362 μM trolox equivalent (TE)/mg. Subsequently, this fraction furtherly purified by Sephadex G-15 chromatography and reversed-phase high performance liquid chromatography (RP-HPLC) showed that the ORAC value was 2.96 ± 0.10 μM TE/mg. The ORAC value of fermented hydrolysates increased from 0.359 μM TE/mg to 0.425 μM TE/mg through 3 kDa ultrafiltration membrane. Five peptides from hydrolysates by enzyme treatment and twelve peptides from fermented scallop mantle hydrolysates were identified using LC-MS/MS. All selected sequences except Lys-Asp-Ala-Ala-Lys-Thr-Lys showed clear differences in scavenging activity for various radicals, which were dependent on the content and position of charged polar amino acids, aromatic amino acids and sulfur-containing amino acid in sequences.