罗非鱼肝脏乙酰胆碱酯酶的双水相萃取

乙酰胆碱酯酶是监测环境中有机磷和氨基甲酸酯类农药的重要靶标物质.应用聚乙二醇(PEG)/磷酸氢二钾(K2HPO4)双水相体系对罗非鱼肝脏乙酰胆碱酯酶的萃取分离进行了研究.采用单因素试验,探讨了PEG1000的质量分数、K2HPO4的质量分数、pH值及NaCl的质量分数对酶萃取的影响,并通过正交试验进一步优化萃取条件.结果表明:K2HPO4质量分数、PEG质量分数、pH值对酶的萃取影响最大.室温下在K2HPO4质量分数为15.0％、PEG1000质量分数为22.0％、pH值为7.0的双水相体系中,酶的分配系数最大、可达5.65,萃取率为72％.放大试验结果表明,该双水相体系对乙酰胆碱酯酶的萃取效果比较稳定,是提取分离乙酰胆碱酯酶的一种新方法.

Extraction of liver acetylcholinesterase in Oreochromis aurea by aqueous two-phase system

Acetylcholinesterase (AChE, EC3.1.1.7) plays an important role in environmental insecticide detection. Extraction of liverAcetylcholinesterase(AChE) in Oreochromis aurea by aqueous two-phase system (ATPS) was studied. The effects of K2HPO4 concentration,polyethylene glycol 1000 (PEG1000) concentration, pH value and NaCl concentration on the extraction of liver AChE were investigated.Orthogonal experiment was used to optimize the extraction of AChE. The results showed that K2HPO4 concentration, PEG1000 concentration and pH value had important effects on partitioning of AChE. Optimum separation of AChE was obtained with a maximumpartition coefficient 5.65 and recovery 72% under the extraction conditions of 15.0% K2HPO4, 22.0% PEG1000 and pH 7.0 at room temperature. Scaled-up experiments indicated this system was stable in separation of AChE.