Preparation, separation and identification of novel hypocholesterolemic peptides from wheat germ: An in vitro and in silico study

The aim of this study was to prepare, isolate, and identify hypocholesterolemic peptides from wheat germ protein and explore their efficacy. Wheat germ protein was hydrolyzed using four commercial enzymes. Hydrolysate, with the highest in vitro hypocholesterolemic activity was isolated using ultrafiltration and macroporous resin. The fractions with highest binding affinity to sodium taurocholate were evaluated for cholesterol-lowering activity and resistance to digestion using Caco-2 monolayers. Fraction III had the highest cholesterol-lowering activity, reducing the subcutaneous transport and absorption of cholesterol and resisted digestion. Nano-LC–MS/MS and molecular docking were used to identify cholesterol-lowering peptides from Fraction III. Three cholesterol-lowering peptides, FAAGAPP, GAGDIPGGIG, and GPVPDTGIFS, were identified. These peptides exhibited cholesterol micelle solubility, specifically by 76.2 %, 68.3 %, and 64.7 %, respectively. In summary, wheat germ peptides exhibited significant cholesterol-lowering activity in vitro, suggesting their potential for application in functional foods. [Display omitted] •WGPs prepared by alcalase showed the best hypocholesterolemic activity.•Fraction III reduced the subcutaneous transport and absorption of cholesterol.•Fraction III possessed excellent resistance to simulated gastrointestinal digestion.•Three novel peptides, FAAGAPP, GAGDIPGGIG, and GPVPDTGIFS, were identified.