Scavenging Activity of Whey Protein Hydrolysates in HaCaT Cells

Abstract

Enzymatic hydrolysis can be an effective tool for improving the cosmeceutical-related properties of a milk protein. As a source of valuable peptides, our study has provided the information for the first time that whey protein hydrolysates could be served as the potential antioxidant active ingredient for alleviating skin aging. The factorial design was used to determine the effect of enzymatic hydrolysis conditions. The studied enzymatic hydrolysis conditions were the types of enzyme (papain, trypsin and chymotrypsin), the enzyme to substrate ratio (1/100, 1/200 and 1/1000), and the hydrolysis time (1, 3 and 5 hrs). Twenty seven whey protein hydrolysates were produced and tested for the anti-oxidative action in human keratinocyte cells. The intracellular reactive oxygen species (ROS) level was determined by a specific oxidative probe DCFH2-DA and flow cytometry. The results indicated that the ROS scavenging activity of such hydrolyzed whey proteins was depended on the types of enzymes and the enzyme to substrate (E/S) ratio. The whey protein hydrolyzed by papain significantly suppressed cellular ROS induced by DMNQ in comparison to those hydrolyzed by trypsin, chymotrypsin, and native whey. The finding thus encourages the utilization and development of these promising products for anti-oxidant and anti-aging approaches.