权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Development of new method for the preparation of enzyme thin films

酶薄膜制备新方法的开发

基本信息

批准号：
09558110
负责人：
ANZAI Jun-ichi
金额：
$ 8.64万
依托单位：
TOHOKU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

项目摘要

Experimental variables for the preparation of enzyme thin films using avidin-biotin technology were investigated in detail. The enzyme thin films containing glucose oxidase (GOx) were found to be used for several months if the GOx films were prepared under the optimum conditions, including the concentration of enzyme solution, temperature, pH, and treatment time. In order to immobilize much amount of enzymes in durable thin films, some kinds of synthetic polymer were employed as material for the film. Biotin-modified polyamines (polyethyleneimine (PEI) and polyallylamine (PAA)) can be used for immobilizing much amount of avidin through avidin-biotin interactions. On the other hand, polyanions such as polyvinylsulfate (PVS) and polystyrenesulfonate (PSS) were useful for immobilize avidin through electrostatic force of attraction, by a layer-by-layer deposition of the polyanion and avidin. The flexible backbone of PVS and PSS is responsible for high loading of avidin in the thin films. In these polyanion/avidin thin films, the biological activity of avidin (or binding ability to biotin) was retained and it was possible to accommodate some biotin-tagged functional molecules in the polyanion/avidin thin films. During the preparation of enzyme thin films by avidin-biotin interactions, it necessary to keep the film wet for the development of well-organized and biologically-active films. If the film was dried, the catalytic activity of enzyme was degraded rapidly, probably due to the degradation of the structure of the film. During the course of this study, we obtained useful information for the development of biologically active enzyme thin films through avidin-biotin interactions.

本文详细研究了用亲和素-生物素技术制备酶薄膜的实验条件。在最佳条件下制备的葡萄糖氧化酶（GOx）膜，包括酶液浓度、温度、pH值和处理时间，可连续使用数月。为了在耐用的薄膜中吸附大量的酶，采用了一些合成聚合物作为薄膜的材料。生物素修饰的多胺（聚乙烯亚胺（PEI）和聚烯丙基胺（PAA））可以通过亲和素-生物素相互作用固定大量的亲和素。另一方面，聚阴离子如聚硫酸乙烯酯（PVS）和聚苯乙烯磺酸酯（PSS）通过聚阴离子和抗生物素蛋白的逐层沉积，通过静电吸引力可用于抗生物素蛋白。PVS和PSS的柔性骨架负责在薄膜中高负载抗生物素蛋白。在这些聚阴离子/亲和素薄膜中，亲和素的生物活性（或结合生物素的能力）被保留，它是可能的容纳一些生物素标记的功能分子在聚阴离子/亲和素薄膜。在利用亲和素-生物素相互作用制备酶薄膜的过程中，为了得到组织良好、具有生物活性的酶膜，必须保持膜的湿润。如果将膜干燥，则酶的催化活性迅速降低，可能是由于膜结构的降解。在本研究过程中，我们获得了有用的信息，通过亲和素-生物素相互作用的生物活性酶薄膜的发展。