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研究生: 李宜芳
Li, I-Fang
論文名稱: 修飾冠狀醚之金奈米粒子對胰蛋白酶活性的影響
The Activity of Trypsin upon Complexation with Thiolalkylated Crown Ether Modified Gold Nanoparticles
指導教授: 葉晨聖
Yeh, Chen-Sheng
學位類別: 碩士
Master
系所名稱: 理學院 - 化學系
Department of Chemistry
論文出版年: 2004
畢業學年度: 92
語文別: 中文
論文頁數: 85
中文關鍵詞: 金奈米粒子胰蛋白脢冠狀醚
外文關鍵詞: Trypsin, Gold nanoparticles, crown ether
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  •   將衍生硫醇基的有機環分子修飾於金奈米粒子上,其有機環分子為兩種冠狀醚,分別為2-(10-硫醇基葵烷氧基)甲基-12-冠狀醚-4和2-(10-硫醇基葵烷氧基)甲基-18-冠狀醚-6,利用電子穿透式顯微鏡(TEM)觀察其修飾冠狀醚的金奈米粒子的互相作用關係。我們發現修飾不同孔洞的冠狀醚於金奈米粒子上,加入磷酸鹽類緩衝液中,其聚集的方式會有所不同,修飾18-冠狀醚-6的金,以最密推積的方式聚集形成二維六角形,而修飾12-冠狀醚-4的金,則是比較偏向三維的雜亂排列。接著,測試金奈米粒子 / 冠狀醚對蛋白脢的影響,固定的酵素濃度均為10μM,分別混合10nM、5nM、1nM等不同濃度的金奈米粒子 / 冠狀醚濃度,並與純胰蛋白酶的酵素活性比較。我們發現在加入濃度為1nM的金奈米粒子 / 冠狀醚的酵素催化之初始速率降低了40%,而當加大金奈米粒子 / 冠狀醚的濃度為10 nM時,初始速率卻與純的胰蛋白酶幾乎相同。

      Two crown ether, 2-[(10-Mercaptodecyl)oxy]methyl-18-crown-6 (18-Crown-6) and 2-[(10-Mercaptodecyl)oxy]methyl-12-crown-4 (12-Crown-4), functionalized gold nanoparticles have been characterized both in its TEM micrograph and its effect on trypsin trytic activity. In TEM micrographic studies, Au/18-crown-6 conjugates organized into a uniform 2D monolayer with close hexagonal packing, while Au/12-crown-4 exhibited both 2D and 3D assembles, in the potassium phosphate buffer (10 mM, pH 8). The trypsin tryptic activity was studied as a function of Au/Crown concentrations (1 nM, 5 nM, 10 nM). It was found that the initial rate in the 1 nM of Au/Crown showed about 40% decrease compared with pure enzyme; however, with the increase of Au/Crown concentration, the activity increased.

    中文摘要 英文摘要 目錄.................................................Ⅰ 表目錄...............................................Ⅳ 圖目錄...............................................Ⅴ 第一章 序論 1.1 奈米材料的介紹................................1 1.2 奈米粒子的性質及種類..........................4 1.2.1 奈米的結構....................................4 1.2.2 表面效應......................................5 1.2.3 結構效應......................................6 1.2.4 量子化效應....................................8 1.2.5 表面電漿共振效應.............................10 1.2.6 膠體粒子之特性...............................13 1.3 奈米粒子的製備方法...........................17 1.4 奈米粒子之表面修飾...........................20 1.4.1 單層自組合薄膜(Self Assembly Monolayers).....20 1.4.2 多層自組裝(Layer by Layer)...................21 1.4.3 巨分子修飾...................................22 1.5 金奈米粒子(Gold nanoparticles)簡介...........24 1.5.1 粒徑 SPR 吸收值及生成方式....................24 1.5.2 製備方法.....................................24 1.5.3 粒徑偵測.....................................26 1.6 胰蛋白酶 (Trypsin) 之功能介紹................30 1.6.1 胰蛋白酶之結構...............................30 1.6.2 胰蛋白酶之酵素活性...........................31 1.7 冠狀醚(Crown ether)之功能介紹................32 1.7.1 冠狀醚的特性.................................33 1.7.2 冠狀醚之常見應用.............................34 第二章 實驗......................................35 2.1 研究動機與目的...............................35 2.2 材料.........................................37 2.3 儀器設備.....................................38 2.4 實驗步驟.....................................40 2.4.1 製備金奈米粒子 (Au nanoparticles)............40 2.4.2 合成冠狀醚衍生物 (Crown thiols)..............41 2.4.3 金奈米粒子表面修飾冠狀醚 (Au/Crown)..........45 2.4.4 量測酵素活性.................................47 第三章 結果與討論................................51 3.1 金奈米粒子......................................51 3.2 金奈米粒子 / 冠狀醚 之特性......................53 3.3 金奈米粒子 / 冠狀醚 / 胰蛋白酶 之特性...........56 3.4 金奈米粒子 / 冠狀醚 / 胰蛋白酶 之酵素活性分析...58 3.6 結論............................................62 參考文獻.............................................80

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