Production of whey protein ingredients with improved bioactivity based on single and sequential enzymatic hydrolysis reactions

Whey protein concentrate (WPC) via single or sequential enzymatic hydrolysis steps using either only Alcalase® 2.4 L (WPH-A) or Alcalase® 2.4 L followed by Flavourzyme® 500 L (WPH-AF) was investigated. Different bioactive properties of particular interest in the obtained hydrolysates were evaluated. Calcium-chelating capacity (Ca-CC) of the hydrolysates was optimized using response surface methodology to adjust the experimental conditions used for producing the calcium nano-compounds. Chelates with higher Ca-CC were obtained using the WPH-AF compared to WPH-A. Additionally, antihypertensive and antioxidant activities of both hydrolysates showed significant improvements when compared to WPC, and these capabilities remained stable after undergoing different heat treatments. Finally, both hydrolysates presented a cytoprotective effect in two cell lines, suggesting that these products may have positive effects against diseases associated with reactive oxygen species. The results obtained indicate that the WPHs produced herein could be used in the development of food formulations with potential health benefits.