摘要： 目前，国内市场上销售的固定化乙酰胆碱酯酶，大多存在稳定性不高、精确度不高、可重复使用性不高等一系列问题。因此本课题着力于研究如何开发一种高效、稳定、灵敏的，能够长期保存在室温下的固定化酶。首先通过乙酰胆碱酯酶的分子模拟对其性质及其对底物的结合性质进行了探究，然后以戊二醛为交联剂完成乙酰胆碱酯酶的固定化，将乙酰胆碱酯酶与载体（NH2-SBA-15）结合，考察不同戊二醛交联浓度（0~20mM）对固定化酶热稳定性的影响，同时通过实验考察了添加不同浓度（0%~0。5%）的添加剂对固定化酶热稳定性的影响，如蔗糖、明胶、牛血清白蛋白等。根据实验结果得出，最佳的戊二醛交联浓度为10mM，且添加0。4%BSA和0。3%明胶时，固定化乙酰胆碱酯酶在室温（25℃）的热稳定性最佳。77502

毕业论文关键词： 乙酰胆碱酯酶；戊二醛；添加剂；热稳定性；分子模拟

Molecular Simulation and Immobilization of Acetylcholinesterase (Thermal Stability Study)

Abstract: Currently, most of the immobilized acetylcholinesterase which the domestic market sales, have not high stability，not high accuracy, not high reusability and a series of problems。 Therefore, this topic focuses on how to develop an efficient, stable, sensitive and long-term immobilized enzyme to preserve at the room temperature。 Firstly, the properties of acetylcholinesterase and the binding properties of the substrate were studied by the molecular modeling of the acetylcholinesterase。 Then the glutaraldehyde as the crosslinking agent immobilized the acetylcholinesterase which made the acetylcholinesterase was incubated with the carrier (NH2-SBA-15)。 The effects of different glutaraldehyde crosslinking concentration (0 ~ 20mM) on the thermostability of immobilized enzyme were investigated。 At the same time, the effects of additives with different concentrations (0% ~ 0。5%), such as sucrose, gelatin and bovine serum albumin on the thermal stability of immobilized enzyme were investigated by experiments, too。 According to the experimental results, the best glutaraldehyde crosslinking concentration was 10mM。 And add 0。4% BSA and 0。3% gelatin, the immobilized acetylcholinesterase at room temperature (25 ℃) had the best thermal stability。

Keywords:Acetylcholinesterase;Glutaraldehyde;Additive;Thermostability;Molecular simulation

目  录

1 研究背景 1

1。1 固定化酶的应用 1

1。1。1 固定化酶基础理论研究的应用 1

1。1。2 固定化酶在化学分析中的应用 1

1。1。3 固定化酶在环境中的应用 2

1。1。4 固定化酶在临床医疗中的应用 2

1。1。5 固定化酶在农残检测上的应用 2

1。2 乙酰胆碱酯酶 3

1。3 乙酰胆碱酯酶固定化 3

1。4 载体材料 4

1。4。1 纳米管 4

1。4。2 磁性高分子微球