Purification of Kallikrein Using Reverse Micellar Extraction / 中国生物工程杂志

ABSTRACT

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 co-surfactant 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/mg to 219U/mg,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.