磁性納米固定化蛋白酶研究

生物工程論文編號(hào):SW004  論文字?jǐn)?shù):11247,頁數(shù):22

目  錄
中文摘要 I
英文摘要 II
目錄 III
1. 緒論 1
 1.1  生物酶催化 1
      1.2  固定化酶的性質(zhì)及其影響因素 1
      1.3  酶固定化方法 3
 1.4 固定化酶與游離酶的比較 5
 1.5 實(shí)驗(yàn)思路   6 
2. 實(shí)驗(yàn)部分 7
 2.1 實(shí)驗(yàn)試劑及配制 7
 2.2 實(shí)驗(yàn)儀器 8
 2.2 酶的選取 8
 2.3 實(shí)驗(yàn)步驟 8
3. 結(jié)果與討論 11
 3.1  實(shí)驗(yàn)結(jié)果  11
 3.2  實(shí)驗(yàn)結(jié)果分析 13
4．總結(jié)與展望 16
致謝 17
參考文獻(xiàn) 18

摘要：酶具有在溫和條件下可以快速催化反應(yīng)等優(yōu)點(diǎn)，但是，在在實(shí)際應(yīng)用中，利用游離酶作為催化劑有很多局限性，因此本文主要研究蛋白酶的固定化。首先用磁性納米材料與3-氨基丙基三乙氧基硅烷相結(jié)合，得到的產(chǎn)物再用戊二醛試劑以共價(jià)鍵與蛋白酶相結(jié)合，得到磁性納米固定化蛋白酶。然后用考馬斯亮藍(lán)染料結(jié)合比色法測(cè)定蛋白酶的固載率。
 實(shí)驗(yàn)結(jié)果得到，用磁性納米鐵可以成功的實(shí)現(xiàn)蛋白酶的固定化。用磁性納米氧化鐵作為載體，蛋白酶的固載率為40.55％，用磁性納米四氧化三鐵作為載體，蛋白酶的固載率為46.35％。因此，磁性納米四氧化三鐵與蛋白酶的固定化結(jié)合較高。
 
關(guān)鍵詞：蛋白酶；磁性納米鐵；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