反胶束萃取阿魏酸酯酶

Purification of Feruloyl Esterase Using Reverse Micelles Extraction

利用十六烷基三甲基溴化铵(CTAB)/正己醇/异辛烷反胶束萃取阿魏酸酯酶.考察pH值、离子强度、CTAB浓度对阿魏酸酯酶的CTAB/正己醇/异辛烷反胶束萃取及其反萃取的影响.结果表明:萃取的最佳条件为酶液透析24h,pH值为12,萃取液为25mmol.L-1 CTAB的正己醇/异辛烷溶液(体积比为1∶5),萃取率接近100%;反萃取的最佳条件为pH值为7的0.20mol.L-1 KCl溶液,反萃取率可达79%.纯化后比活为356.7μkat.g-1,纯化倍数为6.9,SDS-PAGE电泳显示两条带.

The cetrimonium bromide/hexanol/isooetane reverse micelles extraction and backward extraction were used to purify feruloyl esterase. The effects of pH, ionic strength and cetrimonium bromide （CTAB） concentration on the cetrimonium bromide/hexanol/isooetane reverse micelles extraction and backward extraction were studied. The results showed that the extraction efficiency was nearly 100% when the optimal extraction conditions were used as followed： enzyme dialysis time 24 h, pH 12 and CTAB concentration in hexanol/isooctane solution （volume ratio 1 ： 5） 25 mmol· L-1. The optimal condition of backward extraction was 0. 20 mol · L-1 KC1 with pH 7, and the efficiency of backward extraction was 79%. After extraction and backward extraction of the enzyme, specific activity was 356. 7μkat· g-1, purification fold was 6. 9, and SDS-PAGE eleetrophoresis showed two bands.