Purification and identification of antioxidative peptides from mackerel (Pneumatophorus japonicus) protein

Abstract

This study reports the preparation, purification and identification of an antioxidative peptide from mackerel (Pneumatophorus japonicus) protein. Neutrase was chosen as the optimum protease, with the highest cellular antioxidant activity of 53.65%. The optimal hydrolysate conditions for mackerel protein hydrolysates (MPH) according to response surface methodology were an enzyme concentration of 1203.2 U g⁻¹, extraction time of 4.53 h, pH of 7.26, water/material ratio of 5.22 v/w and extraction temperature of 43.72 °C. MPH was separated using ultrafiltration membranes, and the fraction MPH-III with molecular weight below 3500 Da showed the highest cellular antioxidant activity. Five fractions were separated from MPH-III on a Sephadex G-25 column, and MPH-III-2, exhibiting the highest cellular antioxidant activity, was further separated with an XBridge® peptide BEH C18 column. The MPH-III-2-6 separated from RP-HPLC was further analysed by Thermo Scientific Q Exactive mass spectrometer, and the heptapeptide LDIQKEV (843.5 Da) and the octapeptide TAAIVNTA (759.4 Da) were identified. The results of this study offer a promising alternative to produce natural antioxidative peptides from fish protein hydrolysate, which may be utilized as functional ingredients in food systems.