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Abstract Bioactive peptides, produced by the action of buttermilk starter culture or the gastrointestinal enzymes, may have a potential contribution in the health promoting activities of fermented buttermilk. Thus, a comprehensive analysis of buttermilk protein hydrolysate (BMPH), as well as during a simulated gastrointestinal digestion, was performed using ultra performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS). A total of 63, 120 and 46 peptides were identified in fermented buttermilk, gastric and intestinal digests, respectively. Most of these peptides are unique for fermented buttermilk and some of them were previously described as bioactive substances. The main part of bioactive properties was the hypotensive peptides which may partially explain the positive impact of buttermilk on reducing blood pressure in humans. Several antioxidant and hypotensive peptides were produced from the gastric digest hydrolysate (BMGH) of fermented buttermilk. After chemical synthesis of such peptides, the sequence PWDQVKR showed potent antioxidant activity, approximately similar to vit.C, while the peptide SGPLRPFF inhibited the angiotensin converting enzyme (ACE) with an IC50 value as low as 1.56 μM. |