Background: Protein-derived peptide fractions can play a key role in the physiological and metabolic regulation and modulation of the body, which suggests that they could be used as functional ingredients to improve health and to reduce the risk of disease. This work aimed to evaluate the in vitro antithrombotic and anticariogenic bioactivity of hydrolysates and protein fractions obtained from cowpea (Vigna unguiculata) by biocatalysis., Results: Cowpea protein concentrate was hydrolyzed by sequential action with two enzyme systems, Pepsin-Pancreatin or Alcalase-Flavourzyme. There was extensive enzymatic hydrolysis, with degrees of hydrolysis of 34.94% and 81.43% for Pepsin-Pancreatin and Alcalase-Flavourzyme, respectively. The degree of hydrolysis for the control treatments, without the addition of the enzymes Pepsin-Pancreatin and Alcalase-Flavourzyme was 1.1% and 1.2%, respectively. The hydrolysates were subjected to fractionation by ultrafiltration, with five cut-off points according to molecular weight (<1, 1-3, 3-5, 5-10 and >10 kDa). The Alcalase-Flavourzyme hydrolysate led to 100% inhibition of platelet aggregation, while the Pepsin-Pancreatin hydrolysate showed 77.41% inhibition, but this was approximately 100% in the ultrafiltered fractions. The highest anticariogenic activity was obtained with the Pepsin-Pancreatin system, with 61.55% and 56.07% for calcium and phosphorus demineralization, respectively., Conclusion: Hydrolysates and their peptide fractions from Vigna unguiculata exhibited inhibition of platelet aggregation and protection of tooth enamel and have the potential for use in the development of functional products with beneficial health effects. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)