Production and characterisation of whey protein hydrolysate having antioxidant activity from cheese whey

J Sci Food Agric. 2015 Nov;95(14):2908-15. doi: 10.1002/jsfa.7032. Epub 2014 Dec 30.


Background: Cheese whey is a rich by-product in nutritional terms, possessing components with high biological value, excellent functional properties, and an inert flavour profile. In the present study, mozzarella cheese whey was ultra-filtrated to remove lactose and mineral. The retentate was hydrolysed with food-grade enzyme alcalase and the hydrolysis conditions (pH, temperature and time) were optimised for maximum antioxidant activity using response surface methodology.

Results: Whey protein hydrolysed for 8 h at pH 9 and 55 °C showed a maximum antioxidant activity of 1.18 ± 0.015 µmol Trolox mg(-1) protein. The antioxidant peptides were further enriched by ultra-filtration through a 3 kDa membrane. Seven peptides - β-Lg f(123-131), β-Lg f(122-131), β-Lg f(124-131), β-Lg f(123-134), β-Lg f(122-131), β-Lg f(96-100) and β-Lg f(94-100) - were identified by LC-MS/MS in the 3 kDa permeate of the hydrolysate. The incorporation of whey protein hydrolysate (WPH) in lemon whey drink (5-10 g L(-1)) increased the antioxidant activity from 76% to 90% as compared to control.

Conclusion: Hydrolysis of ultra-filtrated retentate of whey can be an energy- and cost-effective method for the direct production of WPH from whey compared to the industrial production of WPH from whey protein concentrate. This study suggests that WPH with good nutritional and biological properties can be effectively used in health-promoting foods as a biofunctional ingredient.

Keywords: alcalase; antioxidant activity; bioactive peptides; cheese whey; response surface methodology; whey protein hydrolysate.

MeSH terms

  • Antioxidants / pharmacology*
  • Beverages
  • Cheese*
  • Citrus
  • Filtration
  • Functional Food
  • Humans
  • Peptides / pharmacology*
  • Protein Hydrolysates / pharmacology*
  • Subtilisins
  • Whey
  • Whey Proteins / pharmacology*


  • Antioxidants
  • Peptides
  • Protein Hydrolysates
  • Whey Proteins
  • Subtilisins