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系統識別號 U0002-1907201115271600
中文論文名稱 應用幾丁質及奈米複合顆粒於幾丁聚醣酶與蛋白酶之純化
英文論文名稱 Application of chitin particles on the purification of chitinolytic enzymes and proteases
校院名稱 淡江大學
系所名稱(中) 化學學系碩士班
系所名稱(英) Department of Chemistry
學年度 99
學期 2
出版年 100
研究生中文姓名 陳政融
研究生英文姓名 Jung-Long Chen
學號 698160594
學位類別 碩士
語文別 中文
口試日期 2011-07-12
論文頁數 50頁
口試委員 指導教授-王三郎
共同指導教授-薛文發
委員-陳佑汲
中文關鍵字 蛋白酶  幾丁質酶  磁性幾丁聚醣/四氧化三鐵納米複合顆粒  粗製蝦殼幾丁質 
英文關鍵字 Protease  chitinase  Magnetic chitosan / Fe3O4 nanocomposite particles  Crude SSC 
學科別分類 學科別自然科學化學
中文摘要 使用幾丁類物質粗製蝦殼粉 (Crude SSC) 與磁性幾丁聚醣/四氧化三鐵納米複合顆粒 (NF) 以吸附方式純化酵素,以達成快速純化之目的。
由吸附劑SP、AC、pure chitin、Crude SSC、Crude CSC、Crude SPC中選擇較適吸附劑Crude SSC。再探討溫度、重量、時間對吸附鳳梨酵素之影響。經由實驗得知於4℃、30分鐘、0.1 g的條件下,得到比活性0.0918 U/mg。
再探討溫度、重量、時間對NF吸附鳳梨酵素之影響。從實驗得知於25℃、30分鐘、0.1 g 的條件,得到比活性0.1357 U/mg。
使用上述條件,使用NF與Crude SSC吸附細菌酵素,藉此討論吸附方式純化酵素的結果。經由實驗可知純化酵素影響條件並非只有等電點的因素,也需考慮酵素和吸附劑的相互作用還有物理吸附等因素。
結果證明吸附方式純化酵素是一個可行的純化方式,可以提高純化效率。
英文摘要 Application of NF and Crude SSC on the purification of enzymes to achieve rapid purification purposes. Adsorbent : SP, AC, pure chitin, Crude SSC, Crude CSC, Crude SPC , Choose a more suitable adsorbent Crude SSC. Studying temperature, weight, time on the adsorption effect of the pineapple enzyme. By experiment that at 4 ℃, 30 min, 0.1 g of the conditions to get specific activity 0.0918 U / mg.
Studying temperature, weight, time on the effect of NF adsorption bromelain enzyme. Learned from the experiment at 25 ℃, 30 min, 0.1 g of the conditions to get specific activity 0.1357 U / mg.
Use the above conditions, NF, and Crude SSC absorption bacterial enzyme purified enzyme to discuss the results. Purified enzyme affected by the experimental conditions known isoelectric point is not the only factor, but also need to consider the interaction between enzyme and adsorbents as well as physical absorption and other factors.
Results show that absorption is a viable way to purify the enzymes, it can improve the purification efficiency.

論文目次 第一章 緒論 1
第二章 文獻回顧 2
2.2 幾丁聚醣之應用 2
2.3蛋白酶 3
2.4幾丁質酶及幾丁聚醣酶 3
2.5吸附劑 4
2.6 吸附劑種類 5
2.7 幾丁質吸附劑 5
2.8磁性幾丁聚醣/四氧化三鐵奈米複合顆粒 (NP) 特性 6
2.9 吸附及純化 6
第三章 材料與方法 8
3.1 實驗材料 8
3.2 實驗儀器 9
3.3 吸附流程 9
3.4 鳳梨酵素的製備 12
3.5 蛋白酶之活性測定 12
3.6 幾丁聚醣酶之活性測定 12
3.7 crude SSC之較適吸附條件之探討 13
3.7.1吸附劑選擇 13
3.7.2 crude SSC吸附濃度 13
3.5.3 crude SSC吸附溫度 14
3.8 NP較適吸附條件之探討 14
3.8.1 顆粒大小之選擇 14
3.8.2 NP添加量之選擇 14
3.8.3 NP吸附溫度 14
3.9 crude SSC和NP於細菌酵素之吸附 15
3.9.1粗酵素液製備 15
3.9.2 粗酵素液之吸附 15
3.9.3 電泳分析 15
第四章 結果與討論 16
4.1吸附劑及吸附條件探討 16
4.1.1 吸附劑之探討 16
4.1.2吸附劑濃度對於吸附鳳梨酵素之影響 17
4.1.3溫度對於吸附之影響 18
4.2 NP之吸附條件探討 20
4.2.1 顆粒大小對於吸附之影響 20
4.2.2 NP添加量於吸附之影響 20
4.2.3溫度對於NP吸附的影響 21
4.2.4時間對於NP吸附之影響 21
4.3 crude SSC與NP吸附細菌酵素之條件探討 23
4.3.1 TKU 011粗酵素液之吸附結果 23
4.3.2 TKU 015粗酵素液之吸附結果 24
4.3.3 TKU 022粗酵素液之吸附結果 24
4.3.4吸附TKU 024粗酵素液之結果 25
4.4細菌酵素吸附之比較 26
第五章 結論 44
參考資料 45

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謝佳霖(2010) Bacills cereus TKU022 所生產幾丁聚醣酶及蛋白酶之特性分析與應用。淡江大學生命科學研究所碩士班碩士論文。
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