Study of screening, transport pathway, and vasodilation mechanisms on angiotensin-I converting enzyme inhibitory peptide from Ulva prolifera proteins

In this study, Ulva prolifera protein was used for preparing angiotensin-I converting enzyme (ACE)-inhibitory peptide via virtual gastrointestinal digestion and in silico screening. Some parameters of the obtained peptide, such as inhibition kinetics, docking mechanism, stability, transport pathway, were explored by Lineweaver-Burk plots, molecular docking, in vitro stimulate gastrointestinal (GI) digestion and Caco-2 cells monolayer model, respectively. Then, a novel anti-ACE peptide LDF (IC₅₀, (1.66 ± 0.34) µmol/L) was screened and synthesized by chemical synthesis. It was a no-competitive inhibitor and its anti-ACE inhibitory effect mainly attributable to four Conventional Hydrogen Bonds and Zn701 interactions. It could keep activity during simulated GI digestion in vitro and was transported by peptide transporter PepT1 and passive-mediated mode. Besides, it could activate Endothelial nitric oxide synthase (eNOS) activity to promote the production of NO and reduce Endothelin-1 (ET-1) secretion induced by Angiotensin II (Ang II) in Human Umbilical Vein Endothelial Cells (HUVECs). Meanwhile, it could promote mice splenocytes proliferation in a concentration-dependent manner. Our study indicated that this peptide was a potential ingredient functioning on vasodilation and enhancing immunity.