酒石酸钾钠双水相体系分离纯化C-藻蓝蛋白

采用廉价的聚乙二醇(PEG)/酒石酸钾钠建立双水相体系,从干螺旋藻中分离纯化藻蓝蛋白C-PC。以纯度(目的蛋白在620 nm处的特征吸光值和总蛋白在280 nm处吸光值的比值)为指标,优化体系参数。研究结果表明,体系p H 8.06,系线长为21.65%、体积比为0.22的PEG1000和酒石酸钾钠双水相体系分离纯化效果最好,纯度由0.42提高到1.27,高于食品级纯度0.7,纯化因子3.04,回收率85.11%。硫酸铵盐析提高粗提液纯度后,二次双水相纯度达到3.28,高于药品级3.0。得到的C-PC具有较高的生物活性。

Separation and purification of C-phycocyanin using aqueous two-phase extraction technique of potassium sodium tartrate

This paper used commercial aqueous two-phase system (ATPS) to efficiently extract and purify C-PC from the dry spirulina platensis. The purity of C-PC was defined as the relation between of 620 and 280 nm absorbance, which was as the target to optimize the system parameters. The optimal conditions were proved in polyethylene glycol (PEG) 1000 and potassium sodium tartrate, system pH of 8.06, the tie-line length (TLL) of 21.65% (w/w) and the volume ratio (Vr) of 0.22 to increase the purity from the initial purity of 0.42 to 1.27 after the first extraction. The purity was over the food grade 0.7 and the purification factor was 3.04. The recovery yield was 85.11%. The purity was achieved up 3.28 after the saturation of ammonium sulfate in the second ATPS extraction. It was more than the pharmaceutical grade 3.0.