Purification of Trypsin by Using AOT/Isooctane Reverse Micellar Extraction and Overcoming Denaturation and Precipitation in the Process
- VernacularTitle:AOT/异辛烷反胶束萃取纯化胰蛋白酶及酶变性失活问题的解决
- Author:
Lin-Jie CHEN
;
Xue-Jun CAO
;
- Publication Type:Journal Article
- Keywords:
Reversed micellar Trypsin Ethanol
- From:
China Biotechnology
2006;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Trypsin was purified from crude material of pig pancreas by AOT/isooctane reversed micellar system.The influence of the main operating parameters such as ethanol concentration in forward and backward extraction,pH,KCl and AOT concentration,temperature were investigated.Forward and backward extraction recovery of trypsin reached almost 90% and neared 100%,respectively.Finally,about 88% of total yield was obtained,and the specific activity of trypsin was increased to over 1800U/mg with purification factor of 5 folds more.In AOT-isooctane reverse micellar extraction system,denaturation or precipitation of proteins always occured due to strong electrostatic interaction between AOT-proteins molecules.It had been resolved by adding ethanol into reverse micellar system,and no denaturation was observed.Otherwise,the phase separation time was shortened significantly because of ethanol added.It was only 10 minutes or less to reach phases separation after forward and backward extraction.If this method can be applied in industry,efficiency will be greatly improved.
