The purpose of this study was to develop an in-vitro digestion protocol to evaluate the antioxidant potential of the peptides found in processed cheddar cheese using digestion enzymes. We first studied antioxidant and angiotensin-converting enzyme (ACE) inhibition and antioxidant activities of processed cheddar cheese with the addition of spices e.g., cumin, clove, and black pepper made from buffalo milk and ripened for 9 months. Then we conducted an in vitro digestion of processed cheddar cheese by gastric and duodenal enzymes. Freeze-dried water (WSE) and ethanol-soluble fractions (ESE) of processed cheddar cheese were also monitored for their ACE inhibition activity and antioxidant activities. In our preliminary experiments, different levels of spices (cumin, clove, and black pepper) were tested into a cheese matrix and only one level 0.2 g/100 g (0.2%) based on cheese weight was considered good after sensory evaluation. Findings of the present study revealed that ACE-inhibitory potential was the highest in processed cheese made from buffalo milk with the addition of 0.2% cumin, clove, and black pepper. A significant increase in ACE-inhibition (%) of processed cheddar cheese, as well as its WSE and ESE, was obtained. Lower IC50 values were found after duodenal phase digestion compared to oral phase digestion.
Keywords: ACE-inhibition; antioxidant potential; ethanol-soluble extract; processed cheddar cheese; water-soluble extract.