基于类水滑石材料固定酶构筑电化学生物传感器
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摘要
生物传感器是一种集微电子学、材料学、生物技术为一体的高新技术产品。它作为一种快速、灵敏的检测技术,正成为食品检测技术研究的新热点。其中基于生物酶构筑的电化学生物传感器的发展在当前生物传感器领域最具代表性。如何选用合适的材料在电极表面有效地固定生物分子,使被固定的生物分子能保持较高的生物活性,并实现对实际样品的检测,对于研制电化学生物传感器具有非常重要的意义,具有广阔的应用前景。
层状纳米材料具有结构规整、化学组成可调变及可插层组装等特性,已成为国内外材料学科研究的热点之一,而且层状纳米材料剥层后的纳米片具有更开放的结构,为酶的固定化带来了新的契机。本论文分别采用层状水滑石纳米材料氯离子插层镁铝水滑石(Mg-Al-Cl LDHs)固定酪氨酸酶(Tyr)、剥离水滑石纳米片(Mg-Al-LDHNS)固定肌红蛋白(Mb),构筑性能优秀的电化学生物传感器。本论文研究内容主要包括以下几个部分：
1、采用氯离子插层镁铝水滑石(Mg-Al-Cl LDHs)作为固定Tyr的载体,Mg-Al-Cl LDHs固定的Tyr保持了良好的生物活性。电化学研究发现,Mg-Al-Cl LDHs固定的Tyr修饰玻碳电极,对儿茶酚实现了良好的电流响应。基于苯甲酸对Tyr活性的抑制作用,实现了对食品中苯甲酸的真样检测。在此基础上,将其成功应用于可一次性使用的丝网印刷电极,有望在食品中的苯甲酸快速检测上得到应用。
2、通过将水滑石剥离重组,合成了Mb-Mg-Al-LDHNS的插层化合物；通过XRD表征,说明Mb-Mg-Al-LDHNS形成了规则的层状结构。大片Mg-Al-LDHNS为Mb提供了适合的分子取向,并实现了Mb与电极之间的直接电子转移。生物分子修饰电极对H2O2保持了催化活力。
Much attention has been devoted to the development of the electrochemical biosensor based on enzymes. It possesses important value in physic, environmental monitoring, food and military. Layered nanomaterials have many specialities, such as orderly structure, adjustable chemical composition, intercalationable, assembliable, and nanosheets obtained from the delamination of layered nanomaterials have a more open structure. Novel electrochemical biosensors have been fabricated based on these layered nanomaterials and nanosheets in this topic. It provides a new idea to develop new type of electrochemical biosensors, and it has broad application prospects.
In this paper, the nanomaterials (layered double hydroxide compounds and exfoliated nanosheets) were used to immobilize Tyrosinase (Tyr) or Myoglobin (Mb) and investigate the electrochemical properties. The main contents of the thesis are given as follows:
1. Mg-Al-Cl layered double hydroxide was applied to the immobilization of Tyr as a support matrix. Tyr almost retained bioactivity in the film. The electrochemical results revealed that it can be applied for determination of food preservative, benzoic acid.The detection of benzoic acid was performed via its inhibiting action on the Tyr/Mg-Al-Cl LDHs modified screen-print electrode. This inhibitor biosensor was successfully applied for the determination of benzoic acid in some real samples.
2. Mb has been intercalated into Mg-Al-LDHNS by delamination and restacking through XRD analysis, and the direct electron transfer between the intercalated Mb and the underlying electrode was studied. Mb intercalated into Mg-Al-LDHNS retained its native secondary structure partly. The Mb-Mg-Al-LDHNS/GCE showed electrocatalytic activity in the reduction of H2O2.
层状纳米材料具有结构规整、化学组成可调变及可插层组装等特性,已成为国内外材料学科研究的热点之一,而且层状纳米材料剥层后的纳米片具有更开放的结构,为酶的固定化带来了新的契机。本论文分别采用层状水滑石纳米材料氯离子插层镁铝水滑石(Mg-Al-Cl LDHs)固定酪氨酸酶(Tyr)、剥离水滑石纳米片(Mg-Al-LDHNS)固定肌红蛋白(Mb),构筑性能优秀的电化学生物传感器。本论文研究内容主要包括以下几个部分：
1、采用氯离子插层镁铝水滑石(Mg-Al-Cl LDHs)作为固定Tyr的载体,Mg-Al-Cl LDHs固定的Tyr保持了良好的生物活性。电化学研究发现,Mg-Al-Cl LDHs固定的Tyr修饰玻碳电极,对儿茶酚实现了良好的电流响应。基于苯甲酸对Tyr活性的抑制作用,实现了对食品中苯甲酸的真样检测。在此基础上,将其成功应用于可一次性使用的丝网印刷电极,有望在食品中的苯甲酸快速检测上得到应用。
2、通过将水滑石剥离重组,合成了Mb-Mg-Al-LDHNS的插层化合物；通过XRD表征,说明Mb-Mg-Al-LDHNS形成了规则的层状结构。大片Mg-Al-LDHNS为Mb提供了适合的分子取向,并实现了Mb与电极之间的直接电子转移。生物分子修饰电极对H2O2保持了催化活力。
Much attention has been devoted to the development of the electrochemical biosensor based on enzymes. It possesses important value in physic, environmental monitoring, food and military. Layered nanomaterials have many specialities, such as orderly structure, adjustable chemical composition, intercalationable, assembliable, and nanosheets obtained from the delamination of layered nanomaterials have a more open structure. Novel electrochemical biosensors have been fabricated based on these layered nanomaterials and nanosheets in this topic. It provides a new idea to develop new type of electrochemical biosensors, and it has broad application prospects.
In this paper, the nanomaterials (layered double hydroxide compounds and exfoliated nanosheets) were used to immobilize Tyrosinase (Tyr) or Myoglobin (Mb) and investigate the electrochemical properties. The main contents of the thesis are given as follows:
1. Mg-Al-Cl layered double hydroxide was applied to the immobilization of Tyr as a support matrix. Tyr almost retained bioactivity in the film. The electrochemical results revealed that it can be applied for determination of food preservative, benzoic acid.The detection of benzoic acid was performed via its inhibiting action on the Tyr/Mg-Al-Cl LDHs modified screen-print electrode. This inhibitor biosensor was successfully applied for the determination of benzoic acid in some real samples.
2. Mb has been intercalated into Mg-Al-LDHNS by delamination and restacking through XRD analysis, and the direct electron transfer between the intercalated Mb and the underlying electrode was studied. Mb intercalated into Mg-Al-LDHNS retained its native secondary structure partly. The Mb-Mg-Al-LDHNS/GCE showed electrocatalytic activity in the reduction of H2O2.
引文
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