Purification and characterization of novel antioxidant peptides of different molecular weights from mackerel Pneumatophorus japonicus protein hydrolysate

Abstract

Mackerel (Pneumatophorus japonicus) proteins were hydrolyzed by five proteases: trypsin, papain, neutrase, acid protease, and flavourzyme. The hydrolysate treated by neutrase exhibited the highest antioxidant activity. Response surface methodology (RSM) was employed to optimize the hydrolysis conditions in an effort to obtain a mackerel protein hydrolysate (MPH) with the highest DPPH radical scavenging activity. The MPH was fractioned using a series of ultrafiltration membranes and five fractions, namely, MPH-I (>10 kDa), MPH-II (10-2.5 kDa), MPH-III (1-2.5 kDa), MPH-IV (0.4-1 kDa), and MPH-V (below 0.4 kDa), were obtained. DPPH radical scavenging activity, reducing power, hydroxyl radical scavenging activity, and the lipid peroxidation inhibition capability of these fractions were evaluated. The fractions in molecular weights <2.5 kDa (MPH-III, MPH-IV, and MPH-V), which occupied 93.4% of the total fractions, showed the strongest antioxidant activity; and the antioxidant activities of the three fractions are similar to each other. Using SP Sephadex C-25 and Sephadex G-25 columns, eight fractions were obtained from the MPH (<2.5 kDa). The isolated peptide I (1 664 kDa) displayed the highest DPPH radical scavenging activity. Therefore, MPH is a potential source of antioxidant peptides.