Abstract:The preparation of AHP from recombinant E.coli BL21 by affinity chromatography and thrombin mediated cleavage was studied through orthogonal experiment. RP-HPLC was used to detect the antihypertensive peptides in AHP. The optimum loading quantity of sample and flow rate were 4 mg (GST-AHP)/mL (medium bed) and 1.5 mL/min, respectively, under which, the absorption capacity of fusion protein was 3.86 mg/mL (medium bed) with a recovery rate of 96.5%. The optimum cleavage conditions of fusion protein by thrombin were as follows: thrombin dosage of 15 U /mg GST, reaction time of 8 h and temperature of 22 ℃, under which 102 μg of AHP could be obtained using 1 mg GST-AHP. In addition, bioactivity of recombinant AHP was studied. The obtained AHP showed high bioactivity in vitro with IC50 and Km of 4.6 μmol/L and 1.95 mmol/L, respectively.
