三維結構具官能基中孔洞材料於酵素固定及染料吸附之應用

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研究生：	巫誠恩 Cheng-En Wu
論文名稱：	三維結構具官能基中孔洞材料於酵素固定及染料吸附之應用 Functionalized Cubic Mesoporous Silica Nanoparticle Materials SBA-1 : Synthesis, Characterization and Applications in Enzyme Immobilization and Selective Dye Adsorption
指導教授：	高憲明 Hsien-Ming Kao
口試委員:
學位類別：	碩士 Master
系所名稱：	理學院 - 化學學系 Department of Chemistry
論文出版年：	2016
畢業學年度：	104
語文別：	中文
論文頁數：	211
中文關鍵詞：	中孔洞材料、蛋白質吸附、選擇性染料吸附
外文關鍵詞：	mesoporous silica, protein adsorption, selective dye adsorption
相關次數：	點閱：6 下載：0
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本論文主要分為兩部分，第一部分是利用陽離子型界面活性Hexadecyl pyridinium chloride (CPC) 以及陰離子型高分子Poly(acrylic acid) (PAA) 作為模板，合成擴孔且具羧酸官能基中孔洞材料，其簡稱為CS-1B-x、MP-CS-1B-x及LP-CS-1B-x，x = [CES/(CES+TEOS)]，並將其應用於固定蛋白質的載體，固定的蛋白質為木瓜蛋白酶 (Papain)。結果顯示，擴孔及羧酸官能基含量提高皆有助於提高吸附量，且含有羧酸官能基之材料蛋白質固定效果較好，活性測試也顯示，吸附木瓜蛋白酶後一系列材料在35o C下也能保有木瓜蛋白酶良好之活性，並且在高溫下可保護木瓜蛋白酶使其不容易失活，並將吸附數據代入等溫吸附模型，發現Langmuir等溫吸附模式較其他等溫吸附模式更適合描述CS-1B-x吸附蛋白酶之系統，動力學則屬於Pseudo-second order之吸附過程，此外亦由分子內擴散模型之速率常數結果，驗證第二步驟之分子內擴散階段為吸附蛋白質之速率決定步驟。
第二部分同樣利用前面合成的CS-1B-0及CS-1B-40作為模板，利用後修飾法植入胺基，合成單純具胺基及同時具羧酸官能基及胺基中孔洞材料，其簡稱為NS-1B-10及CNS-1B-10-10，並將CS-1B-0、CS-1B-40、NS-1B-10及CNS-1B-10-10應用在選擇性染料吸附，實驗結果發現，CNS-1B-10-10在酸性條件下能利用材料中NH3+的官能基團，對陰離子染料Eosin Y (EY) 及 Eosin B (EB) 產生選擇性吸附之效果，在鹼性時能利用材料中COO-的官能基團，對陽離子染料Methylene Blue (MB) 產生選擇性吸附之效果，而CS-1B-40由於帶有羧酸官能基，在pH 3.0到pH 9.0間皆帶負電，因此對陽離子染料Methylene Blue (MB)可以產生選擇性吸附效果。

I have two parts in my study. In my first part, the well-ordered cubic mesoporous silicas CS-1B-x, MP-CS-1B-x, and LP-CS-1B-x (Pm3n) were synthesized successfully via co-condensation of tetraethyl orthosilicate (TEOS) and carboxyethylsilanetriol sodium salt (CES), also functionalized with different ratio of catboxylic groups and enlarge pore by 1,3,5-trimethylbenzene (TMB) under basic conditions, and using Hexadecyl pyridinium chloride (CPC) and Poly(acrylic acid) as template.
The CS-1B-x, MP-CS-1B-x, and LP-CS-1B-x were used as adsorbents for immobilization of papain. These processes were systematically studied by varing time, initial concentration and pH. The LP-CS-1B-40 showed an excellent adsorption capacity for 1138 mg papain per gram adsorbent and low leaching rate .
In my second part, the well-ordered cubic mesoporous silicas CNS-1B-10-10 (Pm3n) were synthesized successfully via co-condensation of tetraethyl orthosilicate (TEOS), (3-Aminopropyl) triethoxysilane (APTES) and carboxyethylsilanetriol sodium salt (CES) under basic conditions, and using Hexadecyl pyridinium chloride (CPC) and Poly(acrylic acid) as template.
The CNS-1B-10-10 were used as adsorbents for selective adsorption of positive and negative charge dyes in different pH. And the results show that CNS-1B-10-10 have well selectivity in acidic and basic condition. In acidic condition, CNS-1B-10-10 selectively adsorb negative dyes owing to NH3+ functional groups. In basic condition, CNS-1B-10-10 selectively adsorb positive dyes owing to COO- functional groups.

中文摘要.....i
Abstract.....ii
謝誌.....iii
目錄.....iv
圖目錄.....ix
表目錄.....xv
第一章 序論.....1
1-1 中孔洞二氧化矽材料.....1
1-1-1 中孔洞材料之沿革.....1
1-1-2 中孔洞的定義.....2
1-2 界面活性劑性質簡介.....4
1-2-1 界面活性劑的種類.....4
1-2-2 微胞的形成與結構.....6
1-2-3 界面活性劑與矽氧化物的交互作用.....8
1-3 官能基化之中孔洞材料.....12
1-3-1 表面修飾官能基之中孔洞材料.....12
1-3-2 表面修飾羧酸官能基之中孔洞材料.....14
1-4 文獻回顧.....16
1-4-1 中孔洞材料SBA-1之合成與介紹.....16
1-4-2 以聚電解質及界面活性劑搭配合成之中孔洞材料.....17
1-4-3 . 中孔洞材料吸附蛋白質之發展及應用.....19
1-4-4 中孔洞材料吸附染料之發展及應用.....26
1-4-5 選擇性吸附染料之發展.....29
1-5 研究動機與目的.....31
第二章 序論.....32
2-1 藥品.....32
2-2 實驗步驟.....35
2-2-1 合成具羧酸官能基的中孔洞SBA-1 (CS-1B-x).....35
2-2-2 合成擴孔且具羧酸官能基的中孔洞SBA-1 (LP-CS-1B-x).....    35
2-2-3 以硝酸溶液裂解孔洞中的模板.....36
2-2-4 蛋白質檢量線之製作.....36
2-2-5 材料CS-1B系列在不同反應時間下之蛋白質吸附實驗.....38
2-2-6 材料CS-1B系列在不同pH值下之蛋白質吸附實驗.....38
2-2-7 材料CS-1B-x在不同初始濃度之蛋白質吸附實驗.....39
2-2-8 溶菌酶活性測試實驗.....39
2-2-9 木瓜蛋白酶活性測試實驗.....40
2-2-10 水解酪蛋白動力學測試實驗.....40
2-2-11 不同溫度下木瓜蛋白酶活性測試實驗.....41
2-2-12 高溫下維持木瓜蛋白酶活性測試實驗.....41
2-2-13 不同pH值下木瓜蛋白酶活性測試實驗.....    42
2-2-14 蛋白質之釋放實驗.....43
2-2-15 選擇性蛋白質吸附 (Papain & Hemoglobin).....43
2-2-16 熱力學吸附實驗.....43
2-2-17 利用後修飾法植入胺基之SBA-1.....44
2-2-18 染料檢量線之製作.....44
2-2-19 四種材料在不同pH下之染料吸附實驗.....46
2-2-20 四種材料在在不同時間下對EY及MB染料吸附實驗.....46
2-2-21 . 四種材料在不同初始濃度對EY及MB染料吸附實驗.....46
2-2-22 四種材料在不同pH下選擇性染料吸附實驗 (MB & EY).....    47
2-2-23 . 四種材料在不同pH下選擇性染料吸附實驗 (MB & EB).....    47
2-2-24 CNS-1B-10-10在不同pH下重複使用性染料吸附實驗.....(MB & EY).....48
2-2-25 熱力學吸附實驗.....48
2-3 實驗設備.....49
2-3-1 實驗合成設備.....49
2-3-2 實驗鑑定儀器.....49
第三章 結果與討論.....51
3-1 CS-1B-x、MP-CS-1B-x及LP-CS-1B-x系列.....51
3-1-1 基本性質鑑定.....51
3-1-1.1 XRD繞射圖譜.....51
3-1-1.2 13C CP/MAS NMR.....54
3-1-1.3 29Si MAS NMR.....56
3-1-1.4 等溫氮氣吸脫附.....58
3-1-1.5 FT-IR 紅外線光譜.....61
3-1-1.6 熱重分析.....63
3-1-1.7 SEM影像.....64
3-1-1.8 TEM影像.....66
3-1-1.9 表面電位.....67
3-1-2 CS-1B-x系列之蛋白質吸附實驗.....70
3-1-2.1 不同反應時間吸附溶菌酶之效果.....71
3-1-2.2 不同初始濃度吸附溶菌酶之效果.....72
3-1-2.3 溶菌酶之活性測試實驗.....73
3-1-2.4 溶菌酶之釋放實驗.....74
3-1-3 中孔洞材料吸附溶菌酶後之性質鑑定.....75
3-1-3.1 等溫吸附模式.....75
3-1-3.2 溶菌酶吸附動力學探討.....85
3-1-3.3 吸附溶菌酶之分子內擴散模型之探討.....90
3-1-3.4 吸附溶菌酶之熱力學吸附探討.....94
3-1-4 擴孔之CS-1B-x系列之木瓜蛋白酶吸附實驗.....96
3-1-4.1 不同反應時間吸附木瓜蛋白質之效果.....96
3-1-4.2 木瓜蛋白酶初始濃度對中孔洞材料吸附之影響.....97
3-1-4.3 不同pH值下吸附木瓜蛋白酶之效果.....98
3-1-4.4 不同孔徑大小之材料對木瓜蛋白酶吸附的影響.....100
3-1-4.5 木瓜蛋白酶之活性測試實驗.....101
3-1-4.6 水解酪蛋白動力學測試實驗.....103
3-1-4.7 不同溫度下木瓜蛋白酶活性測試實驗.....104
3-1-4.8 高溫下維持木瓜蛋白酶活性測試實驗.....105
3-1-4.9 不同pH值下木瓜蛋白酶活性測試實驗.....106
3-1-4.10 木瓜蛋白酶之釋放實驗.....107
3-1-5 中孔洞材料吸附木瓜蛋白酶後之性質鑑定.....108
3-1-5.1 XRD繞射圖譜.....108
3-1-5.2 等溫氮氣吸脫附.....109
3-1-5.3 FT-IR紅外線光譜.....111
3-1-5.4 等溫吸附模式.....113
3-1-5.5 吸附木瓜蛋白酶之動力學探討.....120
3-1-5.6 吸附木瓜蛋白酶之分子內擴散模型之探討.....123
3-1-5.7 吸附木瓜蛋白酶之熱力學吸附探討.....127
3-1-6 選擇性蛋白質吸附 (Papain & Hemoglobin).....128
3-2 CS-1B、NS-1B及CNS-1B系列.....130
3-2-1 基本性質鑑定.....130
3-2-1.1 XRD繞射圖譜.....130
3-2-1.2 13C CP/MAS NMR.....131
3-2-1.3 29Si MAS NMR.....133
3-2-1.4 等溫氮氣吸脫附.....135
3-2-1.5 FT-IR紅外線光譜.....137
3-2-1.6 熱重分析.....139
3-2-1.7 SEM影像.....140
3-2-1.8 TEM影像.....141
3-2-1.9 表面電位.....142
3-2-2 四種材料之染料吸附實驗.....144
3-2-2.1 在不同pH下之8種染料吸附實驗.....144
3-2-2.2 不同吸附時間吸附染料之效果.....149
3-2-2.3 染料初始濃度對中孔洞材料吸附之影響.....150
3-2-2.4 選擇性染料吸附實驗 (MB & EY).....154
3-2-2.5 選擇性染料吸附實驗 (MB & EB).....156
3-2-2.6 重複使用性染料吸附實驗 (MB & EY).....158
3-2-3 中孔洞材料吸附染料之性質鑑定.....159
3-2-3.1 等溫吸附模式.....159
3-2-3.2 四種材料吸附染料之動力學探討.....166
3-2-3.3 吸附染料之分子內擴散模型之探討.....170
3-2-3.4 吸附染料之熱力學吸附探討.....175
第四章 結論.....177
第五章 參考文獻.....178


                                

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