鸡血液中乙酰胆碱酯酶的提取研究

Study on the Extraction of Acetylcholinesterase from Chicken Blood

[目的]为了研究硫酸铵盐析法在乙酰胆碱酯酶分离纯化中的效果,从而利用乙酰胆碱酯酶的活性变化反映农药的污染情况。[方法]采用硫酸铵盐析和等电点区分的方法从鸡血中提取乙酰胆碱酯酶,并探讨其适宜的pH值范围。[结果]结果表明：pH值4.5左右大量的杂蛋白能被除去,可获得较高乙酰胆碱酯酶得率;采用饱和度为50%-60%的硫酸铵盐析效果较好,据此得出较佳的提取工艺。[结论]该研究结果为后续鸡血AChE的提取、分离、纯化等工作提供了相应的参考,同时为利用乙酰胆碱酯酶指示环境受有机磷和氨基甲酸酯类农药污染的后续研究和应用提供了理论依据。

Acetylcholinesterase （ACHE） was extracted from chicken blood by means of salting-out effect using ammonium sulfate as the salting-out agent, so as to make use of changes of the activity of acetylcholinesterase to reflect insecticide pollution situation. The experiment has applied ammonium sulfate salting-out and the isoelectric point differentiation methods to extract acetylcholinesterase from chicken blood and explore the appropriated pH range. The results indicated that most impurities of the protein were removed at pH 4.5 and the good production rate of AChE was obtained. In addition, it was found that 50% - 60% saturated ammonium sulfate could salt out most of impurities. The optimal extraction craft was found and discussed in this study. The resuit of this study has provided corresponding reference for follow-up AchE extraction from chicken blood. Meanwhile, it has provided theoretical basis for the follow-up studies and application of using AchE to indicate the insecticide environment pollution by organophosphorus and carbamates .