In silico identification and experimental validation of two types of angiotensin-converting enzyme (ACE) and xanthine oxidase (XO) milk inhibitory peptides.

Bioactive peptides have received significant attention due to their natural origin, low toxicity, and targeting specificity in the past decade. This study identified highly active ACE/XO inhibitors using molecular simulation and online databases and validated their in vitro antioxidant activity and the mechanisms of molecular interactions. According to computer predictions, Asp-Gly-Gly (DGG) and Asp-Gly-Met (DGGM) were identified as potential hydrolysates of common gastrointestinal peptidases with well water-soluble, non-toxic, and non-allergenic. Fourier transform infrared spectroscopy showed that the two peptides altered the enzyme's secondary structure, decreasing α-helix content by about 13 %, along with increasing β-sheet, randam coli, and β-turns content. Molecular docking and molecular dynamics simulations showed that hydrogen bonding and electrostatic interactions caused DGG and DGGM to form stable and dense complexes with the two enzymes. This study provides a new way for economical and efficient screening of new ACE and XO inhibitory peptides from natural proteins.