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

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(>10kDa),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.