物联网-智慧传输-基于MOFs材料的集成纳米酶的设计合成及其比色传感应用.pdf

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物联网智慧传输基于MOFs材料的集成纳米酶的设计合成及其比色传感应用本文研究了基于MOFs材料的集成纳米酶的设计合成及其比色传感应用，旨在解决传统酶系统的种种缺陷。通过MOFs材料的高效 immobilization，实现了酶的高效固定化，并且MOFs材料的多孔结构和丰富的识别位点可促进酶的固定并防止其泄漏。本文设计了一种基于Fe-PDA材料的INAzymes系统，用于快速检测葡萄糖水平。该系统可以快速检测葡萄糖浓度，检测限为1.1 μM，并且在5.0-100.0 μM葡萄糖浓度范围内具有良好的线性关系。本文设计了一种基于Fe-BTC材料的集成纳米酶系统，用于检测葡萄糖水平。该系统可以快速检测葡萄糖浓度，检测限为2.4 μM，并且在5.0-100 μM葡萄糖浓度范围内具有良好的线性关系。本文设计了一种基于HP-MIL-88B-BA材料的集成纳米酶系统，用于快速检测葡萄糖水平。该系统可以快速响应（10分钟），并且在2-100 µM葡萄糖浓度范围内显示出良好的线性关系，检出限为0.98 µM。本文研究了基于MOFs材料的集成纳米酶的设计合成及其比色传感应用，旨在解决传统酶系统的种种缺陷。该研究为生物传感和临床诊断提供了一种新的策略。关键词：物联网智慧传输，MOFs材料，集成纳米酶，葡萄糖检测，级联反应，色传感。 MOFs材料在酶固定化方面的应用 MOFs材料的高效 immobilization，可以实现酶的高效固定化，并且MOFs材料的多孔结构和丰富的识别位点可促进酶的固定并防止其泄漏。该技术可以解决传统酶系统的种种缺陷，如酶的不稳定性和高成本等问题。 MOFs材料的多孔结构可以提供一个大表面积，利于酶的固定化，并且可以防止酶的泄漏。此外，MOFs材料的丰富的识别位点可以促进酶的固定化，并且可以提高酶的催化活性。基于MOFs材料的集成纳米酶的设计合成基于MOFs材料的集成纳米酶可以通过一种或多种方式来实现，如Fe-PDA、Fe-BTC和HP-MIL-88B-BA等。这些材料可以提供一个高效的酶固定化平台，并且可以实现酶的高效催化反应。集成纳米酶的应用基于MOFs材料的集成纳米酶可以应用于生物传感和临床诊断等领域。该技术可以快速检测葡萄糖水平，并且可以实现高效的酶固定化和催化反应。集成纳米酶的优点基于MOFs材料的集成纳米酶具有多种优点，如高效催化活性、高效固定化、快速检测葡萄糖水平等。该技术可以解决传统酶系统的种种缺陷，并且可以提供一个新型的生物传感策略。未来展望基于MOFs材料的集成纳米酶技术具有很高的应用前景，该技术可以应用于生物传感、临床诊断和工业生产等领域。未来，基于MOFs材料的集成纳米酶技术将会继续发展和完善，并且将会对生物传感和临床诊断等领域产生深远的影响。

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为葡萄糖酸，产生 H

作为中间体。H

与 Fe-PDA 快速反应生成•OH，•OH 进一步氧化

3,3'

，

5,5'-

四甲基联苯胺（

TMB

）。氧化的

TMB

的

UV-Vis

吸光度与葡萄糖浓度成正比，

并且在 5.0-100.0 μM 葡萄糖浓度范围内具有良好的线性关系，检测限为 1.1 μM。INAzymes

系统已成功应用于血糖水平的快速比色检测。该 INAzymes 系统具有高催化活性、出色的

灵敏度和增强的化学稳定性，在临床诊断和生物传感方面具有广阔的前景。

2. Fe-BTC（其中 BTC 为 1,3,5-苯三甲酸）类型的 MOFs 用于构建级联比色测定葡萄

糖的集成系统。MOFs 在扮演（a）HRP 模拟物和（b）作为 GOx 的多孔载体方面发挥双重

作用。

GOx@Fe-BTC

通过一锅法制备。在

GOx

进行酶促氧化过程中生成

的同时消耗

了葡萄糖。在 H

存在下，Fe-BTC 模拟酶催化氧化 TMB，形成蓝色的 OxTMB 产物。

OxTMB 的吸光值（在 UV-Vis 652 nm 处测量）在 5.0–100 μM 葡萄糖浓度范围内线性增加，

检测限为

2.4μM

。

GOx@Fe-BTC

已成功应用于血清中葡萄糖的测定。

3. 具有均匀孔隙率和大表面积的 MOFs 作为固定化酶的载体发挥着重要作用。然而，

诸如响应时间长或酶易泄漏的缺点阻碍了其适用性。在这里，我们报道了硼酸功能化的分

层多孔

MIL-88B

（

HP-MIL-88B-BA

）作为

GOx

的有效固定载体。

HP-MIL-88B-BA

具有分

层的多孔结构和丰富的识别位点，可促进 GOx 固定并防止其泄漏。分层孔提高了底物的传

质效率，从而缩短了响应时间。此外，HP-MIL-88B-BA 可用作 HRP 模拟物。以这种方式，

构建了一种集成纳米酶，用于快速一步检测葡萄糖。

GOx@HP-MIL-88B-BA

对葡萄糖能够

快速响应（10 分钟），并且在 2-100 µM 的范围内显示出良好的线性关系，检出限为 0.98 µM。

该方法提供了一种直接、快速、有效的策略用于合成集成纳米酶并应用于生物传感。

关键词：级联反应；比色传感；葡萄糖; 金属-有机框架；集成纳米酶。

万方数据

III

Design and synthesis of integrated nanozymes based on MOFs

for colorimetric sensing

Postgraduate: Zhenghong Zhao Supervisor: Prof. Fanggui Ye

Major: Analytical Chemistry Research area: Biosensing Grade: 2017

Abstract

Enzymes are powerful biocatalysts and play an important role in industrial processes,

biosensors and biofuel cells. However, the fragility and the relatively narrow application range

severely limit the wide application of enzyme systems. These disadvantages can usually be

overcome by enzyme immobilization. The immobilized enzyme provides a convenient method to

isolate the enzyme from the product and improve the stability of the enzyme, thereby helping to

efficiently recover and reuse expensive enzymes and reduce production costs. Many effective

carriers have been used to immobilize natural enzymes to construct artificial multi-enzyme

systems and have achieved excellent success. However, the natural enzymes are still very

expensive. In recent decades, nanozymes with the characteristics of high stability and low cost,

have become substitutes for natural enzymes. However, the selectivity of most nanozymes is

very limited. By encapsulating natural enzymes and nanozymes in a nanoscale framework, the

design of integrated nanozymes (INAzymes) can overcome the limited selectivity of nanozymes.

Many INAzymes have been developed and used for multiple purposes. Among them,

metal-organic frames (MOFs) are porous crystalline materials with large specific surface area,

uniform cavities and high enzyme encapsulation capacity, and some iron-based / copper-based

MOFs have been proven to have good peroxidase activity. Therefore, MOFs are used to design

and synthesize a variety of INAzymes with excellent properties through adsorption, covalent

bonding, pore encapsulation and co-precipitation. Despite the wide range of applications, there

are still some problems to be solved. For example, zeolite imidazolate frames (ZIF)-base

INAzymes have poor chemical stability; chemical covalent immobilization synthesis method is

cumbersome and enzymes are easily inactivated; micropore MOFs result in low substrate mass

transfer efficiency and long response time. Therefore, it is still meaningful to design more novel

INAzymes with excellent stability, simple synthesis method and high cascade catalytic efficiency.

In this thesis, we use different methods to immobilize glucose oxidase (GOx) in enzyme-like

activity MOFs carriers, design three different INAzymes, and investigate and evaluate their

performance. Based on the cascade reaction, the INAzymes colorimetric sensor detection

platform was constructed, which was applied to the one-step rapid detection of glucose content

万方数据

in serum.

1. ZIF-8 has become one of the most typical examples of nanostructures for multi-enzyme

immobilization due to its economical, mild and easy synthesis process. However, ZIF-8

nanocrystals are easily decomposed under acidic conditions. To solve this problem, the

Fe-polydopamine (Fe-PDA) was bonded with ZIF-8 surface to form ZIF-8@Fe-PDA hybrid

shell with good stability. GOx@ZIF-8 was synthesized by one-pot method, then GOx@ZIF-8

was combined with Fe-PDA to synthesize GOx@ZIF-8@Fe-PDA INAzymes. Fe-PDA not only

prevents acid degradation, but also acts as a simulated HRP. In the INAzymes system, glucose is

converted to gluconic acid by GOx in the presence of oxygen to produce H

as an intermediate.

The H

reacts rapidly with Fe-PDA to generate •OH, which oxidizes

3,3',5,5'-tetramethylbenzidine (TMB). The UV absorbance of oxidized TMB is directly

proportional to the glucose concentration, and has a good linear relationship in the range of

5.0-100.0 μM glucose with detection limit of 1.1 μM. The INAzymes system has been

successfully applied to rapid colorimetric detection of blood glucose levels. The INAzymes

system exhibits high catalytic activity, excellent sensitivity, and enhanced chemical stability,

playing great promise in clinical diagnosis and biosensing.

2. A MOFs of type Fe-BTC (where BTC is 1,3,5-benzenetricarboxylic acid) was utilized to

construct an integrated system for cascade colorimetric determination of glucose. The MOFs

performs a dual function in acting (a) as a horseradish peroxidase (HRP) mimic, and (b) as a

solid support for immobilization of GOx. The MOFs was prepared by a one-pot method. Glucose

is consumed while H

is produced during the enzymatic oxidation by GOx. In the presence of

, the HRP mimic catalytically oxidizes TMB to form a blue-green product. The absorbance

of oxidized TMB (measured at 652 nm, UV-Visible) increases linearly in the of 5.0–100 μM

glucose concentration range, and the detection limit is 2.4 μM. The GOx@Fe-BTC was

successfully applied to the determination of glucose in blood.

3. MOFs of uniform porosity and large surface areas, play an important role as carriers of

immobilized enzymes. However, drawbacks, such as long response times or enzyme leakage

have hindered applicability. Herein, we report on a boronic acid-functionalized hierarchically

porous MIL-88B (HP-MIL-88B-BA) as an efficient immobilization carrier for GOx.

HP-MIL-88B-BA features a hierarchical, porous structure with sufficient recognition sites that

facilitate GOx immobilization and prevent enzyme leakage. The hierarchical porosity increases

the mass transfer efficiency of the substrate, thereby reducing the response time. Moreover,

HP-MIL-88B-BA can be used as a HRP mimic. Based on these factors, an integrated nanozyme

was constructed for rapid one-step detection of glucose. GOx@HP-MIL-88B-BA has a rapid

万方数据

中文摘要

..................................................................................................................................................................

Abstract

...................................................................................................................................................................

III

........................................................................................................................................................................

第一章文献综述

.................................................................................................................................................

1.1

引言

..........................................................................................................................................................

1.2

集成酶

......................................................................................................................................................

1.3

集成纳米酶

..............................................................................................................................................

1.3.1

基于介孔二氧化硅

碳的集成纳米酶

.........................................................................................

1.3.2

水凝胶集成纳米酶

.......................................................................................................................

1.3.3

金属及金属氧化物集成纳米酶

...................................................................................................

1.3.4 MOFs 集成纳米酶

.........................................................................................................................

1.4

总结与展望

..............................................................................................................................................

1.5

论文的选题依据、意义及研究内容

......................................................................................................

参考文献：

...................................................................................................................................................

第二章基于 GOx@ZIF-8@Fe-PDA 的级联反应比色检测血糖

..................................................................

2.1

引言

........................................................................................................................................................

2.2

实验

........................................................................................................................................................

2.2.1

仪器和化学试剂

.........................................................................................................................

2.2.2

表征

.............................................................................................................................................

2.2.3 GOx@ZIF-8@Fe-PDA 的合成

...................................................................................................

2.2.4 GOx@ZIF-8@Fe-PDA 活性测定

...............................................................................................

2.2.5 天然酶活性测定

.........................................................................................................................

2.2.6 动力学研究

.................................................................................................................................

2.2.7 葡萄糖检测

.................................................................................................................................

2.3

结果与讨论

............................................................................................................................................

2.3.1 GOx@ZIF-8@Fe-PDA 的表征

...................................................................................................

2.3.2 GOx@ZIF-8@Fe-PDA 的化学稳定性

.......................................................................................

2.3.3 GOx@ZIF-8@Fe-PDA 的活性

...................................................................................................

2.3.4

催化条件优化

.............................................................................................................................

2.3.5 GOx@ZIF-8@Fe-PDA 的动力学研究

.......................................................................................

2.3.6 GOx@ZIF-8@Fe-PDA 的可行性分析

.......................................................................................

2.3.7

人血清中葡萄糖的检测

.............................................................................................................

2.3.8

储存稳定性和可重复使用性

....................................................................................................

2.4

结论

........................................................................................................................................................

参考文献：

...................................................................................................................................................

第三章新型双功能金属-有机框架用于级联比色法测定葡萄糖

.................................................................

3.1

前言

........................................................................................................................................................

3.2 实验过程

................................................................................................................................................

3.2.1 仪器和化学试剂

.........................................................................................................................

3.2.2 表征

.............................................................................................................................................

3.2.3 GOx@Fe-BTC 的制备

................................................................................................................

万方数据