阴离子交换和羟基磷灰石色谱从螺旋藻中提纯藻胆蛋白的研究

本文采用离子交换层析和羟基磷灰石吸附层析技术从螺旋藻中提取纯化得到藻蓝蛋白和别藻蓝蛋白。通过比较 ＡＮＸ Ｓｅｐｈａｒｏｓｅ ４ Ｆａｓｔ Ｆｌｏｗ（ｈｉｇｈ ｓｕｂ／ｌｏｗ ｓｕｂ）、 ＤＥＡＥ Ｓｅｐｈａｒｏｓｅ Ｆａｓｔ Ｆｌｏｗ和 Ｑ Ｓａｐｈａｒｏｓｅ Ｆａｓｔ Ｆｌｏｗ等阴离子交换树脂的动态吸附容量以及目标产品的纯度，选用 ＤＥＡＥＳｅｐｈａｒｏｓｅ Ｆａｓｔ Ｆｌｏｗ作为层析介质．对离子交换的产物进行了电泳分析，藻蓝蛋白和别藻蓝蛋白的等电点接近，电迁移速率相似。采用羟基磷灰石吸附技术对藻胆蛋白混合物进一步分离纯化，分别得到了藻蓝蛋白和别藻蓝蛋白的纯品，经等电聚焦实验验证显示为均一组成。

ISOLATION AND PURIFICATION OF BILIPROTEINS BY CHROMATOGRAPHY ON ANION ION EXCHANGE AND HYDROXYLAPATITE

Separation of biliproteins from Spirulina platensis extracts was accomplished by ion exchange chromatography on DEAE Sepharose Fast Flow, which appears superior to other three kinds of anion exchangers, i.e., ANX Sepharose 4 Fast Flow (high sub), ANX Sepharose 4 Fast Flow (low sub) and Q Sepharose Fast flow, with respect to dynamic capacity and purity of objective product. The electrophoretic titration curves of c-phycocyanin (CPC) and allophycocyanin (APC) show that these two proteins have similar charge properties and their electrophoretic mobilities are approaching, both of which varies little against pH. Separation of CPC and APC is done by chromatography on hydroxylapatite, which results in products of high purity and homogeneity demonstrated by IEF analysis.