漆酶的逆胶束萃取条件优化的研究.pdfVIP

中国环境科学 2013,33(5) ：904~909 China Environmental Science 漆酶的逆胶束萃取条件优化的研究 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 (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) 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. 关键词： 鼠李糖脂；逆胶束；后萃；漆酶 中图分类号：X17 文献标识码：A 文章编号：1000-6923(2013)05-0904-06 Purification progress of laccase by reversed micelle system. PENG Xin1,2, YUAN Xing-zhong1,2*, HUANG Hua-jun1,2, CUI Kai-long1,2, FANG Zhen-min1,2 ZENG Guang-ming1,2 (1.College of Environmental Science and Engineering, Hunan University, Changsha 410082, China ；2.Key Laboratory of Environmental Biology and Pollution Control, Hunan University, Ministry of Education, Changsha 410082, China). China Environmental Science, 2013,33(5)：904~909 Abstract ：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