摘要:本研究采用多酶分步复合水解方式制备黑豆肽，采用DPPH自由基清除率为评价抗氧化活性的指标，通过超滤、高速逆流色谱、SephadexG-25凝胶过滤色谱等分离纯化技术对黑豆混合肽进行分离纯化，获得抗氧化活性较强的黑豆肽组分。结果表明，大黑豆混合肽的DPPH自由基清除率半抑制浓度（IC50）为62.73mg/mL，小黑豆混合肽的IC50为53.74mg/mL，对抗氧化活性较高的小黑豆肽进行超滤处理后，其IC50为37.74mg/mL，将经过超滤后的肽组分再用高速逆流色谱分离纯化，得到一个肽组分(BSP-HSC)，其IC50为27.91mg/mL。最后将BSP-HSC组分经SephadexG-25凝胶过滤色谱分离得到五个峰，其中P3的抗氧化活性最强，其IC50为25.07mg/mL。

关键词 黑豆蛋白；抗氧化活性肽；分离纯化

Abstract:In this paper, black soybean protein were hydrolyzed with different combinations and steps of three kinds of commercial proteases (the Alcalase protease, Neutrase and Flavourzyme protease)，the DPPH free radical scavenging ability was used as an index. In order to obtain the highest anti-oxidative activity back soybean peptides (BSPs), the ultrafiltration (UF), HSCCC and SephadexG-25 column were used. The results showed that compare with the late-maturing black soybean protein hydrolysates, the early-maturing black soybean protein hydrolysates exhibited better antioxidant ability than the late-maturing one, which with the half inhibitory concentration (IC50)62.73mg/mL and 53.74mg/mL, respectively. And then the early-maturing black soybean protein hydrolysates was further purified using consecutive methods on UF, HSCCC and SephadexG-25 column, and the BSP possessed highest antioxidant capability was got, which with the IC50 25.07mg/mL.  

Keywords  Black soybean protein  Anti-oxidant active peptide  Separation and purification