论文总字数：21862字

摘 要

漆酶（Laccase），属于蓝色多铜氧化酶，具有良好的催化作用，通过对底物的电子提取与转移完成氧化还原反应。漆酶的种类多，对应的底物也很广泛，催化效率与选择性强，能制备成生物传感器对邻苯二酚等酚类物质进行检测。氧化石墨烯具有良好的生物导电性。使用氧化石墨烯修饰电极能大大地提高反应效率和电子转移能力。本实验采用氧化石墨烯，用来修饰玻碳电极，再将漆酶固定到电极上，制备成漆酶生物传感器。并且对漆酶电极的制备过程、电极工作条件进行了改善。对氧化石墨烯修饰的漆酶生物传感器对酚类物质的响应情况、选择性、其稳定性和重现性做了考察。本实验结果得出：最适电解质为醋酸缓冲液；最佳工作电位为0.398 V；缓冲液最适pH为4.5；最适溶剂为DMF；最佳扫描速度为50 mV/s。且其具有较好的抗干扰性、稳定性和重现性，对邻苯二酚有灵敏的响应。

关键词：漆酶 氧化石墨烯 生物传感器 邻苯二酚

The preparation and application of laccase biosensor

Abstrsct

Laccase ,belong to the blue copper oxidase,exists in plants and fungi have good catalytic effect,Through to the substrate complete electronic extraction and transfer REDOX reaction.Variety of laccase, corresponding to the substrate is also very extensive, the catalytic efficiency and selectivity, it can be prepared into biological sensors of catechol phenolics were detected.Oxidative graphene has outstanding electrical conductivity, the specific surface area is big, small particles, strong catalytic effect. The use of graphene oxide modified electrodes can greatly improve the reaction efficiency and electron transfer ability.This experiment to the preparation of graphene oxide, used to modify glassy carbon electrode, then fixed to the electrode, laccase enzyme biosensor preparation into the paint.And the preparation process of laccase electrode and electrode to improve working conditions.Of graphene oxide modified paint enzyme biosensor response of phenolics, selectivity, its stability and repeatability of investigation.The experimental resµLts concluded that the optimal electrolyte for acetic acid solution; Best job in the potential for 0.398mv; The optimal pH buffer 4.5; The optimal solvent of DMF; The best scanning speed of 50 mv/s.And it has good selectivity, stability and repeatability, the catechol with sensitive response.

Keywords: Laccase ;Graphene oxide ;Biosensors;O-dihyoxybenzene

目 录

摘要 Ⅰ

Abstrsct Ⅱ

第一章 文献综述.....................................................................................................1

1.1 生物传感器的简介 1

1.1.1 生物传感器的分类及定义 1

1.1.2 生物传感器的工作原理 2

1.1.3 生物传感器在环境检测中的应用 2

1.2 石墨烯简介 3

1.2.1 石墨烯的基本情况 3

1.2.2 石墨烯的制备方法 4

1.2.3 石墨烯的应用 6

1.3 漆酶的简介 7

1.3.1 漆酶的结构与性质 8

1.3.2 漆酶的催化特征 8

1.3.3 漆酶的生物传感器应用 9

第二章 实验部分 10

2.1实验试剂 10

2.2实验仪器 10

2.3实验步骤 10

2.3.1氧化石墨烯的制备 10

2.3.2石墨烯修饰玻碳电极的制备 11

2.3.3漆酶电极的制备 11

2.3.4酚类物质的电化学行为 11

2.3.5漆酶电极的工作条件优化 12

2.3.6漆酶电极对邻苯二酚响应性能的考察 13

2.3.7漆酶生物传感器的抗干扰性 13

2.3.8电极稳定性和重现性的考察 13

第三章 结果与讨论 14

3.1最适电解质的选择 14

3.2最适缓冲液pH的筛选 14

3.3最适溶剂的选择 15

3.4传感器最适扫描速度的选择 15

3.5最佳工作电位的选择 16

3.6漆酶电极对邻苯二酚的响应 17

3.7漆酶生物传感器的抗干扰性 18

3.8电极的稳定性和重现性 18

第四章 结论与展望 20

4.1结论.................................................................................................................20

4.2展望.................................................................................................................20

参考文献....................................................................................................................22

致谢.............................................................................................................................25

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