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以巢湖水华新鲜蓝藻为处理对象,采用分段盐析法纯化获得了医药级藻蓝蛋白。并通过紫外-可见分光光度法对每一步盐析获得的藻蓝蛋白和杂质液进行光谱扫描,辅助确定每一步盐析(NH_4)_2SO_4的最佳投加量。结果表明,六步盐析中奇数次盐析(NH_4)_2SO_4浓度可选择1.0 mol/L,偶数次盐析(NH_4)_2SO_4浓度可选择1.8 mol/L。光谱扫描进一步发现,一步盐析作用机制在于去除了少量的细胞壁残渣、少量核酸类物质、维生素类和杂蛋白等浅黄色物质,二步盐析能大量去除核酸类和维生素类物质,三步盐析能继续去除少量核酸类和部分其他蛋白质,四步盐析进一步去除其他蛋白对藻蓝蛋白的影响,五步和六步盐析纯化作用并不明显,不能同时将这2种性质相近的蛋白质分离开。所得藻蓝蛋白纯度为3.71,回收率可达45.98%。
To Chaohu algae bloom of fresh cyanobacteria for the treatment of objects, the use of sub-salinity purification obtained medical grade phycocyanin. The spectral scans of phycocyanin and impurity solution obtained from each step of salting out were analyzed by UV-visible spectrophotometry to help determine the optimum dosage of salting-out (NH_4) _2SO_4 for each step. The results showed that 1.0 mol / L NH4Cl2 SO4 can be selected in six - step salting out and 1.8 mol / L NH4SO4 _2 SO4 can be selected in the even - order salting out. Spectral scanning further found that one-step salting out mechanism is to remove a small amount of cell wall residue, a small amount of nucleic acids, vitamins and other proteins and other light yellow substances, two-step salting out to remove nucleic acids and vitamins in large quantities, three salt Analysis can continue to remove a small amount of nucleic acids and some other proteins, four-step salting to further remove the other proteins on the impact of phycocyanin, five and six steps salting purification is not obvious, can not these two similar properties of proteins Separate. The purity of the obtained phycocyanin was 3.71, the recovery rate was 45.98%.