Abstract:

After the enzymatic hydrolysate of porcine femoral collagen prepared by a three-stage hydrolysis procedure
sequentially with alkaline protease, papain and flavourzyme was ultrafiltrated, the fraction with molecular weight below
5 kD, showing an IC50 of 1.362 mg/mL was purified by gel permeation chromatography. The effects of elution solvent,
flow rate and sample loading amount on purification efficiency were examined. The best separation results were achieved
by loading 1% (by volume) of the sample onto the column followed by elution with deionized water at a flow rate of
0.6 mL/min. The eluate fraction with the highest antihypertensive activity, showing an IC50 of 0.401 6 mg/mL, was
harvested. The stability of the antihypertensive peptides was evaluated, and the results demonstrated its good tolerance to
heat, acid and alkali and high solubility at pH 2-6.

Key words: enzymatic hydrolysate of porcine femoral collagen, antihypertensive peptides, gel permeation chromatography, stability, half inhibitory concentration against ACE