The Preparation And Application Of Immobilized Enzyme Reactor Using Two Different Carriers

Background: The immobilization of enzyme through some fundamental immobilization techniques(i.e. non-covalent adsorption, covalent binding, entrapment and encapsulation) could restrict enzyme molecules and keep its catalytic properties. Immobilized enzymes offer several advantages over their free form equivalent, such as better stability, potential for reuse, ease in enzyme and product recovery and continuous operation etc. Therefore, this technology has been widely used in the field of medicine, biology, food industry. The structure and property of carrier material plays an important role in the enzyme activity and immobilization efficiency. In this research, two types of immobilized enzyme reactors are designed and prepared using two different carriers respectively. The main research works were as follows:Chapter 1. The progress of immobilized technologies of enzyme especially the carrier and immobilized methods are well reviewed. Two kinds of immobilized enzymes acetylcholinesterase(ACh E) and Trypsin and their related researches in the thesis are also introduced. The research idea, technology roadmap and innovations of this study are also described in detail.Chapter 2. AChE was immobilized onto the amino magnetic microspheres using glutaraldehyde as crosslinking agent. The effects of various parameters on the immobilization process were systematically investigated. The optimized ACh E enzyme reactor was then characterized using certain physical and chemical method and chromatography. Finally, the immobilized ACh E reactor was used to screen the ACh E inhibitors from the extracts of Huperzia serratum(Thunb.) Trev and to establish a high-throughput ACh E inhibitors screening method.Chapter 3. Trypsin was immobilized on the inner surface of fused silica capillary using glutaraldehyde as crosslinking agent. The immobilized trypsin reactor was combined with capillary electrophoresis technology to build the capillary electrophoresis- trypsin enzyme reactor online digestion and analysis method.Chapter 4. The results are summarized as a whole, and the future of immobilizedenzyme reactor is also proposed in this part.