漆酶的逆胶束萃取条件优化的研究

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.

Purification progress of laccase by reversed micelle system

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 (NaCl, NaBr, KCl 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 KCl (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 further proved that laccase was successfully purified. The research indicated RL-RME has huge potential in extraction industry.