Identification of novel angiotensin I-converting enzyme inhibitory peptides from walnut ( Dode) protein hydrolysates and their vascular endothelial protective properties: and studies.

The objective of this study was to explore novel functional peptides derived from walnut protein isolate (WPI) that exhibited gastrointestinal stability, angiotensin I-converting enzyme (ACE1) inhibitory activity, and endothelial protective properties. Among the 81 detected peptides, 36 peptides with Glu, Leu, Pro, and Asp as the predominant residues were predicted to exhibit ACE1 inhibitory activity. Four promising peptides (TLEPT, TLEPN, VYLEQ, and VLSPQ) were selected for further evaluation based on predicted bioactivity and abundance. assays confirmed significant ACE1 inhibitory activity, with VYLEQ and VLSPQ demonstrating exceptional potency, exhibiting IC values of 27.88 ± 2.45 and 104.75 ± 7.62 μM, respectively. Molecular docking and dynamics simulations further revealed that VYLEQ displayed stronger binding affinity to ACE1 (-8.8 kcal/mol) compared to VLSPQ (-8.4 kcal/mol), facilitated by hydrogen bonding, van der Waals forces, and interactions with Zn at the enzyme's active site, contributing to enhanced stability of the ACE1-VYLEQ complex. Moreover, the EA.hy926 cell assay showed that both peptides promoted nitric oxide (NO) release and inhibited endothelin-1 (ET-1) secretion. Molecular docking revealed that both peptides interact with key proteins that regulate NO and ET-1 mainly through van der Waals forces and hydrogen bonding. These findings suggest their potential benefit in antihypertensive effects. The findings of this study have the potential to increase the economic value of skim walnut meal, and to provide a theoretical foundation for the development of novel functional foods that may have a beneficial effect on the health of patients suffering from hypertension.