A highly active angiotensin I-converting enzyme inhibitory peptide KAKW designed based on the role of C-terminal residue, and its antihypertensive effects on spontaneously hypertensive rats.

Angiotensin converting enzyme (ACE) inhibitory peptide KAKP with IC50 of 7.23 ± 0.30 μM was identified from trypsin hydrolysate of pistachio (Pistacia vera L.) through multiple purifications. A series of peptides were designed based on KAKP to reveal the role of the C-terminal residue in the ACE inhibition, including KAK-Xaa (hydrophobic residue), KAK and KAPK. The designed peptide KAKW exhibited a superior ACE inhibitory effect with IC50 of 2.02 ± 0.20 μM. Molecular docking discovered that the hydrophobic C-terminal residues with ring structure facilitated the interaction with Zn2+ and π-π interactions within the ACE active pocket. Isothermal titration calorimetry revealed that the thermodynamic property of KAKW was enthalpy-driven with a lower Kd (57.7 nM) than KAKP (141 nM). KAKW could decrease 20 mmHg SBP at 4 h and 18 mmHg DBP at 6 h during the acute trial of spontaneously hypertensive rats (SHRs), compared to that Lisinopril dropped 33 mmHg SBP and 28 mmHg DBP at 2 h. After a subacute administration of KAKW for 4 w, the strong inhibition of 61.6 % serum ACE activity and the reduction of 33.3 % angiotensin Ⅱ level were achieved. Although KAKW was less potent than Lisinopril, it provided a more moderate and sustained reduction in blood pressure, beneficial for long-term hypertension management. Meanwhile, KAKW ameliorated gut microbiota dysbiosis in SHRs and increased the levels of fecal short-chain fatty acids, demonstrating advantages over Lisinopril in modulating gut microbiota. This work provides a novel scheme for structure-activity study and a prospective candidate for hypertension management.