Characterization of two novel angiotensin converting enzyme-inhibitory peptides from yellow tuna peptides: Inhibitory mechanism, transport route and network pharmacology analysis.

Angiotensin-converting enzyme inhibitory peptides (ACEIPs) from natural food sources are effective alternatives for treating hypertension. This study aims to screen highly active ACEIPs from yellow tuna peptides, and to elucidate their inhibitory mechanism, stability, transport mechanism and potential antihypertensive mechanism by a multiple approach. Two novel ACEIPs STHPHF (IC50 = 30.71 μg/mL) and LTFSY (IC50 = 20.72 μg/mL) were screened by bio-activity guided approach and de novo sequencing. Inhibition kinetics and molecular docking indicated that STHPHF demonstrated a mixed inhibitor, whereas LTFSY acted as a competitive inhibitor of ACE. STHPHF and LTFSY bound to different active pockets of ACE mainly through hydrogen bonding and hydrophobic interactions. In addition, these two peptides exhibited good stability at 20-100 °C and at pH 2-10, but showed certain activity loss during gastrointestinal digestion, whereas LTFSY has better digestive stability than STHPHF. Caco-2 cells monolayer model assays indicated that LTFSY is mainly absorbed through bypass transport, and possesses an efflux mode related to multidrug-resistant protein. Network pharmacology and molecular docking analysis suggested that LTFSY regulates hypertension by multiple pathway and multi-target mode, AGTR1, MAPK8, NOS3, and RHOA maybe the major targets. This study highlights the promising potential of peptide LTFSY for lowering blood pressure.