凝胶层析法分离猪股骨降血压肽及其体外稳定性

Purification of Antihypertensive Peptides from Porcine Femoral Collagen by Gel Permeation Chromatography and Their Stability in vitro

将猪股骨酶解液超滤分离后,对分子质量小于5 k D的酶解滤液(IC50值为1.362 mg/m L)进行凝胶层析分离,研究洗脱液、流速、上样量3个因素对猪股骨降血压肽分离纯化效果的影响。结果表明:以去离子水为洗脱液,流速0.6 m L/min,上样量体积分数1%时,能够得到最佳分离效果,其最高活性组分IC50值达到0.401 6 mg/m L。研究凝胶层析分离后抑制率最高的组分的稳定性,发现其有较好的耐热和耐酸碱、耐盐性,在p H 2~6中有良好的溶解性。

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 k D, showing an IC50 of 1.362 mg/m L 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 m L/min. The eluate fraction with the highest antihypertensive activity, showing an IC50 of 0.401 6 mg/m L, 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 p H 2-6.