摘要:本研究采用多酶分步復合水解方式制備黑豆肽，采用DPPH自由基清除率為評價抗氧化活性的指標，通過超濾、高速逆流色譜、SephadexG-25凝膠過濾色譜等分離純化技術對黑豆混合肽進行分離純化，獲得抗氧化活性較強的黑豆肽組分。結果表明，大黑豆混合肽的DPPH自由基清除率半抑制濃度（IC50）為62.73mg/mL，小黑豆混合肽的IC50為53.74mg/mL，對抗氧化活性較高的小黑豆肽進行超濾處理后，其IC50為37.74mg/mL，將經過超濾后的肽組分再用高速逆流色譜分離純化，得到一個肽組分(BSP-HSC)，其IC50為27.91mg/mL。最后將BSP-HSC組分經SephadexG-25凝膠過濾色譜分離得到五個峰，其中P3的抗氧化活性最強，其IC50為25.07mg/mL。

關鍵詞 黑豆蛋白；抗氧化活性肽；分離純化

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