兼具双重识别和抗粘附功能的无酶型葡萄糖电化学传感器构建及其在微生物发酵过程监控中的应用基础

In fermentation systems, glucose not only serves as the energy source for microorganisms but also is an important raw material and industrial product, and such that monitoring its dynamic content changes is of great significance for the control of microbial metabolisms, identification of production conditions, management of product qualities, and warning of production accidents. In this project, cheap transition metal-based porous electrodes are first utilized as a sensing matrix, and both molecular imprinting and derivative boronic acid-double hydroxyl recognition sites are then introduced to fabricate enzyme-free electrochemical sensors with anti-adhesion and double-recognition properties, to finally realize the high-performance detection of glucose in microbial fermentation environments. In this project, the design and construction of nonenzymatic glucose electrochemical sensors for bio-fermentation process monitoring will be regarded as the main line, and the microbial interception of the sensing interface and its molecularly specific recognition will act as the critical points. The anti-adhesion property against microorganisms will be well regulated via controlling the interface pore size, and the combination of molecularly imprinted sites and boronate-based affinities will be employed to improve the detection selectivity. Based on this research, the interception mechanism of interface pore structures against microorganisms and their structure-activity relationship will be clarified, and the underlying mechanism and regulation rule of the synergistically molecular recognition enhancement between imprinting sites and boronate-based affinities will be revealed, and finally, a new method will be established for the detection of the given target in complex systems with highly selective recognition and favorable stability.

在发酵体系中，葡萄糖既为微生物提供能量来源，又是重要的生产原料和工业产品。掌握其含量的动态变化对调控微生物新陈代谢、辨识生产状况、管理产品质量和预警生产事故至关重要。本项目拟以具有微生物拦截作用的廉价过渡金属基多孔电极作为传感基体，进而同时引入分子印迹和衍生化硼酸-双羟基识别位点，构建兼具抗粘附和双重识别功能的无酶电化学传感器，实现对微生物发酵环境中葡萄糖的高性能检测。项目以设计和构筑无酶型葡萄糖电化学传感器用于生物发酵过程的监控为主线，以传感界面的微生物拦截和分子特异性识别为关键控制点，通过调控界面孔尺寸实现抗微生物粘附，联合非共价分子印迹与共价硼亲和的双重识别能力提高检测选择性。在此基础上，阐明界面孔结构对微生物的拦截机制和构效关系，揭示印迹位点与硼亲和协同增强葡萄糖分子识别的作用机理与调控规律，建立复杂体系中目标物的高特异性识别和高稳定性分析的新方法。

葡萄糖为微生物提供能量来源，也是疾病诊断的重要监测指标。掌握其含量的动态变化对调控微生物新陈代谢、管理产品质量，疾病诊断和控制至关重要。如何经济有效地监控葡萄糖含量是食品安全与控制、临床诊断等领域的研究热点和难点。本项目在执行期间，以多孔镍泡沫作为多孔传感基体，利用水热法制备的碳点，电沉积法制备的过渡金属基中空材料作为电极修饰材料制备修饰电极传感基体。在此基础上，结合原位聚合法、电聚合法、溶胶-凝胶法等分子印迹技术，研制了系列对发酵样品及血样中葡萄糖具有高吸附容量和高选择性的分子印迹传感器。在对所制备材料的表面化学组成、形态等研究基础上，考察了材料的表面结构和化学性质对传感器的电化学性能的影响，探索了分子印迹传感器对发酵样品及血样中葡萄糖的选择性识别性能。结果表明，所构筑的基于硼亲和的双重识别型电化学传感器用于葡萄糖的无酶检测，具有相对较宽的线性范围（0.005～8 mM）及较低的检测限（1 μM）（S/N=3）。所构筑的葡萄糖分子印迹传感器中，对葡萄糖检测的最低检测限可达4.6 nM（S/N=3）。本项目所构筑的传感器具有优异的选择性、较高的灵敏度及良好的再生性能，在食品安全和控制、临床诊断等领域李的均有潜在的经济和社会效益。