Angiotensin-Converting Enzyme (ACE) plays a key role in the blood pressure regulation system.ACE was purified from pig lung by optimized salt fractionation, ion exchange chromatography and ultrafiltration, and its enzymatic properties were studied.The effect of reduced glutathione (GSH) on ACE activity was investigated by using the food-derived ACE inhibitory peptide Ala-Gly-Pro (AGP) as a control inhibitor.The recovery of ACE activity of salt fractionation (the protein mass concentration of pig lung homogenate was 30 mg/mL) and ion exchange chromatography (at 35 ℃) were 75.6% and 61.9%, respectively.Then the final purity (475-fold) was achieved by ultrafiltration and generated a specific activity of 1.9 U/mg and at 43.4% recovery of ACE activity.The enzyme loss during isolation and purification processes was reduced without increasing the cost, and the purification efficiency was improved.The molecular mass of the purified ACE determined from SDS-PAGE analysis was 160 kDa.Moreover, the enzyme was optimally active at pH 7.5 and 37 ℃ with Km value of 2.05 mmol/L and Vmax of 4.64 nmol/L.In vitro activity test indicated that both AGP and GSH exhibited the ACE (from pig lung) inhibitory activity.Moreover, AGP was a competitive ACE inhibitor (IC50 values of 564.0 μmol/L) and GSH was a non-competitive ACE inhibitor (IC50 values of 26.2 μmol/L).This study provides a possibility for screening effective antihypertensive peptides from antioxidant substances for the prevention of hypertension.
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