In the present study, corn protein hydrolysate (CPH) with antioxidant activity was obtained by enzymatic hydrolysis. Corn gluten meal (CGM) was hydrolyzed using two proteases (Alcalase and Protamex) to produce the antioxidant peptide. Extrusion and starch removal of corn protein were used as pretreatment procedures before proteolysis. Hydrolysis by Alcalase has more remarkable digesting efficiency on corn protein than that by Protamex. Therefore, the hydrolysate catalyzed by Alcalase was fractionated by ultrafiltration, and peptide with the highest antioxidant activity was purified from <6 kDa molecular weight fraction. The amino acid sequence of the novel peptide was Gln-Gln-Pro-Gln-Pro-Trp as identified by a quadrupole time-of-flight mass spectrometer (Q-TOF2), with molecular weight of 782.34 Da, which was matched to gamma-zein f (50-55). The new peptide was further synthesized by Fmoc solid-phase method. It showed scavenging activity against DPPH, ABTS, and hydroxyl free radicals in dose dependent manner with EC50 values of 0.95, 0.0112 and 4.43 mg/mL, respectively. It also exhibited notable reducing power of 0.54 at 2.0 mg/mL, but showed weaker Fe2+-chelating capacity with EC50 value of 6.27 mg/mL. These results suggest that the hexapeptide is a potential natural antioxidant that can be used as drug or functional food ingredient. (C) 2014 Elsevier Ltd. All rights reserved.