Structure identification and inhibitory mechanism evaluation of three novel angiotensin converting enzyme (ACE) inhibitory peptides from small-aroma chicken

Small-aroma chicken with a high protein content, is widely distributed in the mountainous area of Yunnan-Guizhou Plateau, China. In this work, three novel ACE-inhibitory peptides, Leu-Thr-Glu-Lys-Val-Val-Phe (LTQKVVF), Leu-Asp-Asp-His-Phe-Leu (LDDHFL), and Val-Pro-Gly-Pro-Glu-Pro-Lys-Pro (VPGPEPKP), were prepared from the Small-Aroma Chicken breast. The purification and identification protocol employed comprised ion-exchange chromatography, gel filtration chromatography, reverse phase-high performance liquid chromatography (RP-HPLC), and nano liquid chromatography-electrospray ionization–tandem mass (Nano-LC-ESI–MS/MS). Compared with VPGPEPKP and VPGPEPKP, LTQKVVF displayed noticeable ACE inhibitory activity, with IC50 values of 189.34 μM. What's more, the Lineweaver-Burk plots revealed peptides LTQKVVF and LDDHFL acted as the mixed-competitive inhibitor while VPGPEPKP inhibited ACE in a manner similar to uncompetitive inhibition. The binding free energies of LTQKVVF, LDDHFL, and VPGPEPKP for ACE were −8.1 kcal/mol, −8.6 kcal/mol, and −9.6 kcal/mol, respectively. The strong inhibition of ACE by VPGPEPKP may be attributed to the interaction of hydrogen bonds, alkyl, and pi-alkyl, which formed by crucial residues of ACE.