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以螺旋藻为原料,在反复冻融、均质和超声联用的基础上,比较了盐提和水提两种不同方式的蛋白提取率,并以水提螺旋藻蛋白为原料,利用胰蛋白酶、胰凝乳蛋白酶、胃蛋白酶、碱性蛋白酶和木瓜蛋白酶分别进行单酶解、双酶解以及体外模拟胃肠道消化的三酶解,得到6种酶解肽。水解度及脱色前后的抗肿瘤活性鉴定结果表明,水解度与抗肿瘤活性无直接相关性,而脱色会降低酶解组分的抗肿瘤活性。对5种活性显著的酶解肽进行超滤,得到9个对乳腺癌和肝癌细胞株的体外生长均有显著抑制作用的组分,并对其中的碱性蛋白酶0~3 K组分进行Sephadex G-15分离纯化,得到对MCF-7和Hep G-2均有较强抑制作用的组分A3,其IC50值分别为64.59μg/m L和61.65μg/m L;得到对Hep G-2具有较强选择性抑制作用的组分A1,A4和A5,其IC50值分别为47.67μg/m L、79.44μg/m L和69.54μg/m L。本研究成果对以螺旋藻为原料的药品和保健食品的开发提供了理论基础。
Spirulina as a raw material, repeated freezing and thawing, homogenization and ultrasonic based on the comparison of salt extraction and water extraction of two different ways of protein extraction rate, and water to extract spirulina protein as raw material, the use of trypsin , Chymotrypsin, pepsin, alkaline protease and papain were single digestion, double digestion and digestion in vitro gastrointestinal digestion of the three enzymatic hydrolysis, six kinds of enzymatic peptide. The degree of hydrolysis and the anti-tumor activity before and after decolorization showed that there was no direct correlation between the degree of hydrolysis and anti-tumor activity, while the decolorization would reduce the anti-tumor activity of the enzymolysis component. Five of the five highly active peptides were ultrafiltrated to obtain nine components that had significant inhibitory effects on the growth of breast cancer and hepatocellular carcinoma cell lines in vitro. The fractions of 0-3 K of alkaline protease were subjected to Sephadex G-15 was isolated and purified to obtain a component A3 with a strong inhibitory effect on MCF-7 and Hep G-2 with IC50 values of 64.59 μg / ml and 61.65 μg / ml, respectively. Hep G-2 The components A1, A4 and A5 with stronger and selective inhibition have IC50 values of 47.67μg / ml, 79.44μg / ml and 69.54μg / ml, respectively. The results of this study provide a theoretical basis for the development of pharmaceuticals and health foods based on spirulina.