1.Beijing Key Laboratory of Food Processing and Safety, Beijing Forestry University, Beijing, 100083, China ;
Abstract:
Angiotensin converting enzyme (ACE) is considered as main causative agent in growing hypertension and other cardiovascular disorders. Inhibition of ACE by producing and purifying bioactive peptides of fermented goat milk is aimed in this study. Protein extracted from goat milk was hydrolyzed with proteolytic enzymes of LH (Lactobacillus helveticus-cicc22171). ACE inhibitory peptides were purified from fermented samples of goat milk protein by optimizing incubation time to 8 h (S-8), 16 h (S-16), 24 h (S-24) and 36 h (S-36), via ultrafiltration. S-8 was used as control to compare the ACE inhibition trend. Molecular weight cut-off; 10000 Da (PM-10) and Ultracel 3K membrane was used to perform ultrafiltration. Sample with 24 h incubation time was considered as best hydrolyzed as compared to others, by applying Nin-Hydrin reaction and SDS-PAGE analysis. ACE inhibitory assay validated the authenticity of S-24 in inhibiting ACE, in vitro. Furthermore, Q executive hybrid quadrupole-orbitrap mass spectrometry was used to determine molecular structure and amino acid sequence of ACE inhibitory peptides. Three peptides, VLPVPQKAVPQ, VLPVPQKVVPQ and TQTPVVVPPFLQPEIMGVPKVKE containing functional amino acid structure, has been identified with highest ACE inhibitory activity on the basis of intensity, size and higher concentration of hydrophobic amino acids as shown in figure as graphical abstract. Fermented goat milk containing these novel bioactive peptides, can be used as nutraceuticals to inhibit ACE and control hypertension in future.