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HepG-2均有较强抑制作用的组分A3，其IC50值分别为64.59 μg/mL和61.65 μg/mL；得到对HepG-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47.67 μg/mL、 79.44 μg/mL和69.54 μg/mL。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。

Based on the combination of repeated freeze-thaw cycles, homogenization, and ultrasonic-extraction, the extraction rate between salt-extraction and water-extraction of Spirulina proteins was compared. Taking water-extracted Spirulina proteins as the raw material, six types of Spirulina proteolytic peptides were obtained through enzymolysis of Spirulina proteins using trypsin, chymotrypsin, pepsin, alkaline protease, and papain individually, using alkaline protease and papain together, and using trypsin, pepsin, and chymotrypsin to simulate the digestive processes of the gastrointestinal tract in vitro. Results of the degree of hydrolysis and anti-proliferation effects before and after decolorization suggested that there was no direct correlation between the degree of hydrolysis and the anti-proliferation effects. In addition, decolorization was found to lower the anti-tumor activity of hydrolytic components. Therefore, five types of proteolytic peptides with good anti-proliferation effects were used for ultrafiltration, providing nine components that could significantly inhibit the growth of both MCF-7 and HepG-2 in vitro. Then, the 0~3 K component of alkaline protease was isolated and purified using a Sephadex G-15 column to obtain the A3 component, which had a strong anti-proliferation effect on MCF-7 and HepG-2 cells, showing IC50 values of 64.59 μg/mL and 61.65 μg/mL, respectively. Additionally, another three components, A1, A4, and A5, were obtained, which had strong selective anti-proliferation effects on HepG-2 cells, with IC50 values of 47.67 μg/mL, 79.44 μg/mL, and 69.54 μg/mL, respectively. These results provide a theoretical basis for the development of new drugs and health foods using Spirulina platensis as raw materials.

国家863项目子课题（2014AA022004）