Purification and Identification of ACE Inhibitory Peptides from Porcine Skin Collagen
Article
Figures
Metrics
Preview PDF
Reference
Related
Cited by
Materials
Abstract:
After being pretreated by ultrasound combined with dilute alkali, porcine skin underwent enzymatic hydrolysis for 30 min to produce hydrolysate with ACE inhibitory activity. This hydrolysate was used as raw material, and the collagen ACE inhibitory peptides were separated and purified using Sephadex G-15 semi-preparative high performance liquid chromatography. The results showed that the optimum separation conditions were as follows: sample concentration of 100 mg / mL; injection volume of 2 mL; flow rate of 2 mL / min; distilled water as the eluent; and a 1 m × 10 mm chromatography column. Under these separation conditions, the mixed peptides were divided into two fractions, AP-I and AP-II. The half maximal inhibitory concentration (IC50) values were 19.50 mg/mL and 1.01 mg/mL, for AP-I and AP-II, respectively. Fraction AP-II, with relatively strong ACE inhibitory activity, was further separated by semi-preparative high performance liquid chromatography, and eight peaks were obtained. The IC50 values of peaks two, five, and six were the lowest (0.44 mg / mL, 0.4 mg / mL, and 0.73 mg / mL, respectively). Based on the IC50 value and the weight of the samples collected from semi-preparative high performance liquid chromatographic separation, liquid chromatography tandem mass spectrometry sequence analysis was conducted for peaks two and six. The results showed that the peptide sequences of peaks two and six were QGPPGPAGPR and AGPPGPPGPA, respectively. Additionally, these sequences occupied amino acid positions 526-535 and 730-739 of the collagen α1 chain, respectively.
