改性与修饰壳聚糖固定化酶纯化抑肽酶研究  

A New Method of Preparation of High purity Aprotininum by Chemical Modified Trypsin of Chitosan .

作　　者：宋扬[1] 侯司 赵辉[1] 王荣海[1] 吴中华[1] 

机构地区：[1]安徽省生物研究所,合肥230031

出　　处：《生物化学与生物物理进展》2000年第1期82-86,共5页Progress In Biochemistry and Biophysics

基　　金：安徽省"九五"科技攻关计划项目!(961 30 1 4 )

摘　　要：采用化学改性与修饰微珠壳聚糖为载体,共价法偶联牛胰蛋白酶,制成抑肽酶亲和吸附剂,单位活力5190KIU/g(湿),蛋白质偶联率605%,酶活性回收率55%;将其直接亲和层析牛肺提取液,分离纯化高比活抑肽酶.方法过程简单,样品比活力5700KIU/mg,质量稳定,成本较低;该吸附剂机械强度高,抗污染能力较强,非特异性吸附较小,可以反复使用,价格低廉,适合工业化生产.The trypsin was covalently linked with chemical modified granulechitosan and was used to isolate and purify aprotininum from the extract of cattle lungs by affinity chromatography. Then high purity aprotininum was prepared after ultrafiltrating and freeze drying. The result showed:the specific activity of immobilized trypsin on chitosan was 25 950 kU/g ,60 5%protein was coupled,and the activity reccvery of trypsin was 55%. The purity of aprotininum was high,and the activity reccvery of trypsin on immobilized trypsin had low non specific adsoption and ideal anti contemination,and it could be used more than 72 times. It was accepted as a simple and stable method and suitable for purifying aprotininum with high activity in industrial manufacturing .

关 键 词：固定化酶 抑肽酶 壳聚糖 化学改性 修饰 

分 类 号：Q81[生物学—生物工程]