Virtual screening and directional preparation of xanthine oxidase inhibitory peptides derived from hemp seed protein

Abstract

The traditional nutritional and medical hemp (<em>Cannabis sativa L.</em>) seed protein were explored for the discovery and directional preparation of new xanthine oxidase inhibitory (XOI) peptides by structure-based virtual screening, compound synthesis, <em>in vitro </em>bioassay and proteolysis. Six subtypes of hemp seed edestin and albumin were <em>in silico</em> hydrolyzed by 29 proteases, and 192 encrypted bioactive peptides were screened out. Six peptides showed to be XOI peptides, of which four (about 67%) were released by elastase hydrolysis. The peptide DDNPRRFY displayed the highest XOI activity IC₅₀ = (2.10 ± 0.06) mg/mL, acting as a mixed inhibitor. The pancreatic elastase directionally prepared XOI hemp seed protein hydrolysates, from which six high-abundance XOI peptides encrypted three virtually-screened ones including the DDNPRRFY. The novel outstanding hemp seed protein-derived XOI peptides and their virtual screening and directed preparation methods provide a promising and applicable approach to conveniently and efficiently explore food-derived bioactive peptides.