摘要:本研究通过碱提酸沉法提取黑豆蛋白，采用多酶分步协同水解黑豆蛋白，用超滤、高速逆流色谱以及凝胶过滤色谱对黑豆蛋白混合肽进行分离纯化，以胆固醇胶束溶解度抑制率为评价指标，测定黑豆蛋白混合肽及其分离纯化组分的降胆固醇活性，筛选降胆固醇活性较强的黑豆肽组分。

    黑豆肽通过超滤系统后，其胆固醇胶束溶解度抑制率由20.58%增大到26.57%，然后采用高速逆流色谱分离纯化，收集120-140min内洗脱下来的组分，抑制率为44.10%。降胆固醇肽组分经凝胶过滤色谱分离得到五个峰，测定分离后各峰的胆固醇胶束溶解度抑制率，得出产生第三个峰的组分对胆固醇胶束溶解度的抑制活性最强，抑制率为50.47%，其相对分子质量为453-916之间。

关键字 黑豆蛋白；降胆固醇活性；分离纯化

Abstract:This study by alkali and acid extraction of black soybean protein precipitation method,using multiple enzyme step hydrolysis of black soybean protein,soybean protein on mixed peptides were separated and purified by ultrafiltration,high-speed countercurrent chromatography and gel filtration chromatography,cholesterol micellar solubility inhibition rate as the evaluation index,the determination of the cholesterol-lowering activity of black bean protein component of hybrid peptide and its purification,screening cholesterol-lowering activity of black beans peptide component.

    Black beans peptide by ultrafiltration system,cholesterol micellar solubility inhibition rate of 20.58% increase to 26.57%,and then using the high-speed counter-current chromatography separation and purification,collection of components eluted in the 120-140min,inhibition rate was 44.10%.Cholesterol-lowering peptide component by gel filtration chromatography,five peaks of separation cholesterol micellar solubility measured after separation of each peak inhibition rate obtained generating component of the third peak is the strongest inhibitory activity on micellar solubility of cholesterol,inhibition rate was 50.47%,and the relative molecular mass of between 453-916.

Keywords  Black bean protein  Cholesterol activity  Separation and purification