反胶束萃取技术分离胰激肽原酶  

Purification of Kallikrein Using Reverse Micellar Extraction

作　　者：王金枝[1] 曹学君[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室生物工程系,上海200237

出　　处：《中国生物工程杂志》2007年第3期93-99,共7页China Biotechnology

摘　　要：研究了用十六烷基三甲基溴化铵(CTAB)/正己醇/正辛烷反胶束溶液萃取和反萃取商业用胰激肽原酶时,水相pH值、离子强度和种类、CTAB浓度和助表面活性剂浓度等因素对分离效率的影响,并从反胶束微观结构给予解释。结果表明:[CTAB]=0.02mol/L,正己醇/正辛烷(V/V)=1∶5,萃取pH=9.0,反萃pH=7.0,萃取[KBr]=0.1mol/L,反萃[KBr]=1.5mol/L,反萃取加15%乙醇(V/V)时,萃取率接近100%,反萃取活性回收得率在80%以上。商业用酶的纯化倍数最高为1.97倍,粗酶为7.15倍,且粗酶纯化后比活在200U/mg以上,电泳分析证实了纯化效果,显示了很好的工业前景。The purification of kallikrein using CTAB/hexanol/octane reverse micelles extraction has been studied and optimized, under various aqueous pH values, ionic strength and species, CTAB concentration and cosurfactant concentration. The result shows that the extraction efficiency approaches 100%, and the activity recovery is more than 80%, the commercial enzyme specific activity is increased by 1.97 times and the crude enzyme activity is increased by 7.15 times, which from 31U/rag to 219U/rag,under the conditions of[ CTAB] = 0.02 mol/L, hexanol/octane （V/V） = 1 ： 5, pH = 9.0 and [ KBr] = 0.1 mol/L in forward extraction, pH = 7.0 , [ KBr ] = 1.5 mol/L and 15% ethanol （V/V） in backward extraction. The result of purified kallikrein is examined by the SDS-PAGE analysis. Reverse micelles extraction is a potential technique for the application in the downstream biotechnological processes.

关 键 词：胰激肽原酶 反胶束 萃取 反萃取 CTAB 

分 类 号：Q814.1[生物学—生物工程]