磁性纳米固定化蛋白酶研究

生物工程论文编号:SW004  论文字数:11247,页数:22

目  录
中文摘要 I
英文摘要 II
目录 III
1. 绪论 1
 1.1  生物酶催化 1
      1.2  固定化酶的性质及其影响因素 1
      1.3  酶固定化方法 3
 1.4 固定化酶与游离酶的比较 5
 1.5 实验思路   6 
2. 实验部分 7
 2.1 实验试剂及配制 7
 2.2 实验仪器 8
 2.2 酶的选取 8
 2.3 实验步骤 8
3. 结果与讨论 11
 3.1  实验结果  11
 3.2  实验结果分析 13
4．总结与展望 16
致谢 17
参考文献 18

摘要：酶具有在温和条件下可以快速催化反应等优点，但是，在在实际应用中，利用游离酶作为催化剂有很多局限性，因此本文主要研究蛋白酶的固定化。首先用磁性纳米材料与3-氨基丙基三乙氧基硅烷相结合，得到的产物再用戊二醛试剂以共价键与蛋白酶相结合，得到磁性纳米固定化蛋白酶。然后用考马斯亮蓝染料结合比色法测定蛋白酶的固载率。
 实验结果得到，用磁性纳米铁可以成功的实现蛋白酶的固定化。用磁性纳米氧化铁作为载体，蛋白酶的固载率为40.55％，用磁性纳米四氧化三铁作为载体，蛋白酶的固载率为46.35％。因此，磁性纳米四氧化三铁与蛋白酶的固定化结合较高。
 
关键词：蛋白酶；磁性纳米铁；固定化

Study of Immobilization of Protease On Magnetic Nanoparticles

Abstract:The Enzyme showed the advantage of rapid reaction in mild conditions ,however, there are many limitations in application of free enzyme. So in this paper，immobilization of protease was studied. Superparamagnetic nanoparticles were first immobilized with 3-aminopropyltriethoxysilane. Then glutaraldehyde was employed as a linker to the protease to the surface of nanoparticles to form Iron Oxide-protease. And then the immobilization yield of protease is measured with Bradford.
 The results show that protease is immobilized successfully with maghemite nanocrystals. Under the condition of the maghemite nanocrystals Fe2O3 as a vector , the immobilization yield of protease is 40.55 percent, and under the condition of the maghemite nanocrystals Fe3O4 as the vector, the immobilization yield of protease is 46.35 percent. So, the immobilization of maghemite nanocrystals Fe3O4 and protease is better.
 
Keywords: protease；maghemite nanocrystals；immobilized