乳酪蛋白源ACE抑制肽的分离纯化

Separation and purification of angiotensin converting enzyme inhibitory casein peptide

本研究以酪蛋白为原料,采用微生物蛋白酶A水解,其酶解产物对血管紧张素转化酶(ACE)的活性有较强的抑制作用(水解度17.4%)。采用DA201-C型大孔树脂脱盐,样品脱盐率达到85%以上,ACE抑制率提高一倍,并用SephadexG-15(层析柱规格1.6cm×85cmI.D.)对脱盐后产物分离纯化,洗脱液为0.02moL/L醋酸缓冲液;反相高效液相色谱(色谱柱μBondapakC183.9mm×300mmI.D.和μBondapakC1819mm×300mmI.D.)在以流动相A梯度洗脱的条件下对产物进一步分离纯化,经纯度分析最终得到单一纯品,其体外ACE抑制率达到84.4%和79.6%。

ACE-inhibitory peptides have been produced by the enzymatic hydrolysis of milk proteins（casein）,One inhibitory peptide from the hydrolyzate at a degree of hydrolysis of 17.4% was purified and identified .It was first isolated by macroporous adsorption resins then isolated by size exclusion chromatography（SEC）, eluted with 0.02 moL/L acetic acid solution on a Sephadex G-15 column（1.6cm×85cm LD,）.The peak of No.5 on the SEC chromatography was further purified by reversed-phase FIPLC （μBondapak C18 3.9 mm×300 mm LD.and μBondapak C18 19mm×300mm LD.）, eluted with a linear gradient elution with acetonitrile from 0 to 50% at room temperature . Then the pure peptide with ACE inhibitory activity was obtained. And ACE inhibitory activities are 84.4% and 79.6%.