油菜蜂花粉ACE抑制活性水解物的制备及分离

Preparation and Separation of ACE Inhibitory Hydrolysates from Rape Bee Pollen

采用4种蛋白酶水解油菜蜂花粉蛋白制备ACE抑制活性物质,高效液相色谱法测定油菜蜂花粉蛋白水解物对ACE的抑制率。结果表明:油菜蜂花粉蛋白酶水解物具有ACE抑制活性,水解物对ACE的抑制活性差异显著(P<0.05),其中碱性蛋白酶>中性蛋白酶>木瓜蛋白酶>酸性蛋白酶,碱性蛋白酶水解物的IC50为0.35mg/mL。4种蛋白酶水解物经BioP-2凝胶分离后,ACE抑制活性较强的组分主要集中在保留时间70～120min,在此区间碱性蛋白酶水解物分离组分对ACE的抑制率达到90%以上,分子质量在376.4～1355D之间。

Four kinds of proteases were used for rape bee pollen protein enzymolysis to prepare ACE inhibitory activity hydrolysates.The ACE inhibitory activity（IC50） of enzymatic hydrolysates was determined by HPLC method.The results indicated that four hydrolysates prepared with different proteases had significantly different ACE inhibitory activities（P 0.05）.The strongest ACE inhibitory activity was observed in alkaline protease derived hydrolysate,with an IC50 of 0.35 mg/mL,followed by neutral protease,papain and acid protease derived hydrolysates.Furthermore,four rape bee pollen protein hydrolysates were applied to a Bio P-2 gel column for fractionization.Fractions with a retention time ranging from 70 to 120 min presented stronger ACE inhibitory activity and statistical analysis showed that their ACE inhibition inhibitory ratios were significantly different（P0.05）.All of the corresponding fractions separated from alkaline protease derived hydrolysate had more than 90% ACE inhibitory ratio,and their molecular weight distribution varied from 376.4 to 1355 D.