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Application of a Novel Synthesized Azo-Dye Ligand for the Affinity Separation of Alkaline Phosphatase
Author(s): 
Pages: 563-567
Year: Issue:  6
Journal: JOURNAL OF CHEMICAL ENGINEERING OF CHINESE UNIVERSITIES

Keyword:  亲和层析染料配基偶氮染料碱性磷酸酶;
Abstract: 设计合成了一种含有碱性磷酸酶抑制剂--对氨基苯磷酸的新型偶氮染料配基并将其偶联到凝胶Sepharose CL-6B 上,制备出新型的亲和层析介质,分离纯化小牛肠中的碱性磷酸酶.通过考察不同配基密度的层析介质对碱性磷酸酶的选择性,得出配基密度为4.55mg 配基.(g 湿胶)-1的层析介质选择性高.用NaCl和Na2HPO4进行阶段洗脱,可使酶的纯化倍数一步达到65倍,活力回收率达到89%.通过测定不同配基浓度下酶的动力学,发现新合成的配基是碱性磷酸酶的竞争性抑制剂,其KI为3.0×10-3mol.L-1.
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