【作者】 管仁峰；

【导师】 张歆；

【作者基本信息】 汕头大学 ， 应用化学， 2006， 硕士

【摘要】 固定化酶作为一种生物催化剂，它能在较为温和的反应条件下，高选择性、高效率地催化某些化学反应，并且不会对环境造成污染。酶的固定化是最具发展前景的生物技术前沿领域之一，在酶反应装置、生物燃料电池以及生物传感器中有着相当大的应用价值。目前，固定化技术主要有吸附法、共价键合法、物理包埋法和交联法等。本文综述了介孔材料的合成、表征及改性，用不同的方法制备了具有大孔径氨基功能化介孔SiO₂和氨基功能化介孔SiO₂膜，并研究了它们在生物分子固定化中的应用。以碳纳米粒子固定化酶，研究了它们在生物燃料电池方面的应用。 主要研究工作如下： 1．我们用微乳模板法制备了直径在17～34nm的介孔二氧化硅（LMCFs）。氨基功能团是通过TEOS在介孔二氧化硅上水解经共价健结合在孔道的内外表面。材料的结构以及化学性质经TEM，XRD，FT—IR和氮气吸附来表征。葡萄糖氧化酶的固定化是用戊二醛共价健连接材料和酶分子中的氨基。固定化的酶仍具有相当高的活性，并在热稳定性方面优于游离酶。固定化的酶可以通过简单的离心分离而被重复利用。 2．用溶胶—凝胶法制备了有序的—NH₂功能化介孔SiO₂膜，用TEM、AFM、FT—IR和激光共聚焦显微镜对膜进行了表征。DNA通过共价键与膜表面的—NH₂结合。介孔膜有序的介孔结构、表面高密度的—NH₂功能化基团大大提高了DNA分子在基体上的固定质量和密度。用溴化乙锭作为荧光增强剂，通过荧光强度表征了吸附于表面的DNA的相对密度。 3．我们制作了一个比较新型的生物燃料电池，电池的两个电极分别用葡萄糖氧化酶和漆酶。葡萄糖氧化酶被固定在碳纳米粒子修饰的阳极，氧化葡萄糖。漆酶用同样的方法固定在阴极，和氧气反应。通过检测电池的性能可证明用碳纳米颗粒修饰的酶电极能促进葡萄糖氧化酶和漆酶的直接电化学。更多还原

【Abstract】 Immobilized enzyme is a type of biocatalyst, which can high-efficiently, high-selectively catalyze chemical reactions under mild conditions and doesn’t pollute the environment. Enzyme immobilization is considered as one of very promising biotechnology, immobilized enzyme is of considerable interest for their applications as enzyme reactors, biological fuel cells and bio-sensors. Immobilization technology includes absorption, wrapping, covalently binding, and crosslinking methods, and so on. In this thesis, the synthesis, characterization, improment were summarized. Besides, ammo-functionalized mesostructured SiO₂ （AF-MCFs） materials and amino-functionalized mesostructured SiO₂ film were synthesized. Their application was also described. We used carbon nanoparticles to immobilize enzyme and fabricate biological fuel cells. The performance was measured. The main research work is as follows:1. Large mesopores cellular foam （LMCFs） materials with diameters ranging from 17 to 34 nm were synthesized using microemulsion templating. The amine functional groups were attached to channels of LMCFs materials via post-synthesis grafting. The structural and chemical properties of these prepared materials were characterized by TEM, XRD, FT-IR and nitrogen adsorption. The glucose oxidase （GOx） was immobilized by covalently couple enzyme molecules to the interior surface of amino-functionalized mesostructured cellular foams （AF-MCFs） materials, in which leaching of the enzyme is prevented. The immobilized enzyme exhibited the high catalytic activity and thermal stability for oxidation of glucose. It was found that GOx immobilized on AF-MCFs materials is re-useabie.2. Ordered amino-functionalized mesoporous organic-inorganic hybrid silica thin films were prepared by evaporation induced self-assembly （EISA） at acidic condition, and DNA molecules were covalently bound to these mesoporous substrates. These prepared materials were characterized by TEM. AFM. FT-IR and Laser Scanning Confocal Fluorescence Microscopy （LSCFM）. The ordered mesoporous structure and highly dense covalent bonds help to increase the combining intensity and DNA density.3. We report the fabrication of a novel biofuel cell containing a glucose oxidase anode and a更多还原