漆酶的逆胶束萃取条件优化的研究  

Purification progress of laccase by reversed micelle system

作　　者：彭馨[1,2] 袁兴中[1,2] 黄华军[1,2] 崔凯龙[1,2] 方振敏[1,2] 曾光明[1,2] 

机构地区：[1]湖南大学环境科学与工程学院,湖南长沙410082 [2]湖南大学环境生物与控制教育部重点实验室,湖南长沙410082

出　　处：《中国环境科学》2013年第5期904-909,共6页China Environmental Science

基　　金：国家自然科学基金资助项目(50978087/21276069)

摘　　要：研究了生物表面活性剂逆胶束体系萃取纯化漆酶的后萃的过程.在此逆胶束体系中,表面活性剂采用一种阴离子生物表面活性剂鼠李糖脂(rhamnolipid,RL),溶剂采用异辛烷.分别考察了4种不同离子种类(NaCl,NaBr,KCl和KBr)与不同浓度、4种不同助溶剂种类(正丁醇,正己醇,正庚醇及正辛醇)与不同比例、乙醇的含量以及温度对漆酶的后萃过程的影响.结果表明,0.25mol/L KCl或0.20mol/L KBr能够得到较好的萃取效果;在四种助溶剂中,正己醇(比例为0.5)和正庚醇(比例为0.5)效果比较好;乙醇加入量为9%效果最优;最适宜温度为35℃.在上述条件下,漆酶的酶活回收率和纯化倍数分别高达90%,4.7倍.电泳分析结果进一步证实了漆酶的纯化效果较好.研究证明鼠李糖脂逆胶束萃取酶类具有广阔的应用前景.Backward extraction of laccase by an innovative method, biosurfactant-based reversed micellar extraction （RME） was studied. In this reversed micelle system, rhamnolipid （RL）, a kind of anionic biosurfactant, was employed and isooctane was used as solvent. Four different salts type （NaC1, NaBr, KC1 and KBr） with different concentration and four different co-solvent type（n-butanol, n-hexanol, n-heptanol and n-octanol） with different ratio were added respectively in the RME. The effects of ethanol and temperature on backward extraction were studied. The experimental results showed that the impact of KCI （0.25mol/L） on backward extraction approximately equal to KBr （0.20mol/L）. Optimum co-solvent was n-hexanol or n-heptanol both with the ratio of 50%. Ethanol was added in backward extraction process and the appropriate volume was 9%. The optimum temperature during backward was 35℃. The best activity recovery and purification fold were 90% and 4.7. The analysis results of SDS-PAGE fi.trther proved that laccase was successfully purified. The research indicated RL-RME has huge potential in extraction industry.

关 键 词：鼠李糖脂 逆胶束 后萃 漆酶 

分 类 号：X17[环境科学与工程—环境科学]