化学改性与修饰壳聚糖固定化酶制备高比活抑肽酶新工艺  

A New Method for Preparation of High - Purity Aprotinin by Chemical Modified Trypsin on Chitosan

作　　者：宋扬[1] 赵辉[1] 侯司 袁克文 石华峰 汪姗霖 

机构地区：[1]安徽省生物研究所,合肥230031 [2]安徽省阜阳生化制药厂

出　　处：《中国生化药物杂志》1999年第4期180-182,共3页Chinese Journal of Biochemical Pharmaceutics

基　　金：安徽省"九五"科技攻关项目!(项目编号9613014)

摘　　要：目的 研究一步层析制备高比活抑肽酶新工艺。方法 将化学改性与修饰壳聚糖体，共价偶联配基牛胰蛋白酶，制成固定化酶为抑肽酶亲和吸附剂，直接对黄牛肺提取液一步层析纯化，再超滤脱盐、浓缩、去热原，冻干即抑肽酶。结果 抑肽酶比活５７００ｋｉｕ／ｍｇ，收率２２×１０＾４ｋｉｕ／ｋｇ（牛肺）；固定化酶单位活力２５９５０ｋｉｕ／ｇ（干）。酶活性回收率５５％，非特异性吸附较小，机械强度高，可反复使用。结论Purpose: Research and establish a 'one-step' technology for preparation of aprotinin with high purity and high specific activity. Methods: Covalent coupling bovine trypsin with chemically modified chitosan, a kind of immobilized enzyme was obtained and used as the matrix of affinity chromatography for preparation of aprotinin from crude extract of bovine lung directly. To condense the sample and remove the salts and pyrogen, the elute was treated continuously by super- filtration, and finally , lyophilized aprotinin was achieved by freeze-drying. Results: The unit activity of immobilized enzyme matrix was 25 950 kin/Gram (dry weight), and the matrix has less nonspecific absorption, much more times of repeated utilization and hight operation pressure. The specific activity of the final product of aprotinin was as high as 5 700 kin/mg, the recovery rate of aprotinin activity was 55%, and 220 000 kiu aprotinin activity could be obtained from one kilogram bovine lung. Conclusion: The lowcost 'one-step' technology for preparation of aprotinin with high purity and high specific activity is suitable for lager scale industrial production.

关 键 词：固定化酶 抑肽酶 壳聚糖 化学改性 化学修饰 

分 类 号：Q556.9[生物学—生物化学] TQ925[轻工技术与工程—发酵工程]