Isolation, virtual screening, action mechanisms, chelation with zinc ions, and stability of ACE-inhibitory peptides from ginkgo seed globulin

Abstract

Ginkgo seed has potential applications in the prevention and treatment of hypertension, but its application in food is limited. Thus, ginkgo seed globulin was hydrolyzed using dual enzymes (Alcalase and thermolysin). After gel column separation, reverse-phase high-performance liquid chromatographic purification, and ESI-MS/MS analysis, five oligopeptides containing fewer than 12 amino acid residues were obtained. Among them, the heptapeptide Glu–Ala–Ser–Pro–Lys–Pro–Val (EASPKPV) offered relatively high capacities to inhibit ACE (IC₅₀: 87.66 μmol L⁻¹) and bind with zinc ions (5.35 ± 0.32 mg g⁻¹). Moreover, EASPKPV showed competitive inhibitory kinetics against ACE. Fourier-transform infrared spectroscopy analysis evidenced that the amino group and carboxyl group of EASPKPV could both provide binding sites for zinc ions. EASPKPV can restrain ACE in the following ways: (i) competitively linking with five key residues (Gln281, Ala354, Glu376, Lys511, and Tyr523) in the S1 and S2 pockets of ACE by short hydrogen bonds; (ii) binding to thirteen active residues of ACE via hydrophobic interactions; and (iii) binding with residue His383 or the zinc ion of zinc tetrahedral coordination. Additionally, simulated gastrointestinal digestion did not show any remarkable efficacy on the capacities of EASPKPV to restrain ACE and bind with zinc ions. These results indicate that ginkgo peptides may be used for antihypertension.