磺胺甲基异噁唑作配基的高效亲和色谱分离胰凝乳蛋白酶和胰蛋白酶

Separation of Chymotrypsin and Trypsin by High-performance Affinity Chromatography with Sulfamethoxazole Ligand

用磺胺甲基异噁唑作配基、大孔硅胶为基质的高效亲和色谱分离纯化胰凝乳蛋白酶及胰蛋白酶。20多种蛋白质和酶在该柱上无特异性吸附。磺胺甲基异噁唑在硅胶上的键合量为4．3mg／g；该亲和填料对胰凝乳蛋白酶吸附量为9．6mg／g；两者之间的亲和解离常数K1／M为1×103mol／L。

Sulfamethoxazole (SMZ ) was reportcd as an affinity ligancl for the separation and Purification of chymotrypsin and trypsin. It was found that both enzymes can interact specifically with SMZ immobilized on large pore silicagel while more than twenty Proteins and enzymes show no interaction with immobilized SMZ. The capacity of the affinity inatrix forchymotrypsin is 9. 6 mg/g. The affinity dissociation constant K1/M, between immobilized SMZand chymotrypsin in solution is 1 ×10 6 mol/L which is located in the optimal range of the dissociation constant from 10 4 to 10 8 mol/L,.