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研究生:鍾建中
研究生(外文):Chung-Chen Chung
論文名稱:利用酵素降解幾丁聚醣之動力學研究
論文名稱(外文):Degradation Kinetic of Chitosan using Enzyme
指導教授:吳和生吳和生引用關係
指導教授(外文):Ho-Shing Wu
學位類別:碩士
校院名稱:元智大學
系所名稱:化學工程學系
學門:工程學門
學類:化學工程學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:61
中文關鍵詞:幾丁聚醣葡萄糖胺木瓜蛋白酵素鳳梨蛋白酵素降解反應
外文關鍵詞:chitosanglucosaminepapain(form papaya latex)bromelain(from pineapple stem)degradation reaction
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摘要
近年來,葡萄糖胺在治療退化性關節炎這方面相當具有療效,而葡萄糖胺的相關產品漸漸的開始普及,葡萄糖胺的產品療效與藥物性質,很少討論葡萄糖胺的製造流程與方式,故本研究主要是利用生物性的酵素降解幾丁聚醣,利用酵素加速降解反應,討論製備葡萄糖胺的可行性以及降解反應在動力學上的探討。目前是利用呈色法與HPLC(High Performance Liquid Chromatography)的來檢測是否在降解反應後會產生葡萄糖胺,經過這2種方法檢測後,可以確定含有葡萄糖胺,再以測量粘度的方式探討幾丁聚醣在不同種類酵素(bromelain、papain、lipase等)、不同酵素含量、不同轉速以及不同溫度下降解反應之動力學探討。
在葡萄糖胺的檢測這方面利用了2種方法(呈色法與HPLC法)可以確定Papain(form papaya latex)降解反應出來的產物中含有葡萄糖胺,而Bromelain(from pineapple stem)雖然會對幾丁聚醣的黏度迅速的減少,但無法在HPLC的測試中得到應有的圖譜,所以目前無法證明幾丁聚醣在Bromelain(from pineapple stem)的反應下,會產生葡萄糖胺。
經過DEAE(diethylaminoethyl) column分離純化後的Papain(form papaya latex)與Bromelain(from pineapple stem),可以確定Papain(form papaya latex)與Bromelain(from pineapple stem)並不是單一的蛋白質酵素,經純化分離後所得的結果,這2種蛋白質酵素Papain(form papaya latex)與Bromelain(from pineapple stem)裡面含有2種成分的蛋白質。
關鍵字:幾丁聚醣、葡萄糖胺、木瓜蛋白酵素、鳳梨蛋白酵素、降解反應。
Abstract
Nearly years, glucosamine salts have good curative effect in arthritis. Then glucosamine products have begin available to all. When I compare references that about glucosamine products, most references almost refer to curative effect of product and property of medicine, but discussed in process and how to made glucosamine that is very deficient. So this research that mainly confer to use bio-enzyme degraded chitosan and increase degradation of reaction. We focus on made up glucosamine discuss the research feasibility and discuss kinetics of degradation. Now we used color test and HPLC(High Performance Liquid Chromatography) that 2 methods measure glucosamine in degradation of chitosan. After this 2 kind of determination, we can confirm the glucosamine exist in residual degradation of chitosan, and used to Ubbelohde capillary viscometer, and measured time (ηt ) at 25℃ in circular bath. By measuring time (ηt ), we discussed the kinetics in different conditions(different ).
In this research,we make sure Papain(form papaya latex) that can produce glucosamine in degradation reaction of chitosan by proving to color test and HPLC determination. Bromelain (from pineapple stem) cause to decrease viscosity of chitosan in the reaction, but bromelain (from pineapple stem) have no peak in HPLC determination. Now, we can’t make sure bromelain (from pineapple stem) that can produce glucosamine in degradation reaction of chitosan.After DEAE (diethylaminoethyl) column separating papain(form papaya latex) and bromelain(from pineapple stem), we can make sure that they are not single protein of enzyme. Papain(form papaya latex) and bromelain(from pineapple stem) both have 2 components of the protein.
Key word: chitosan, glucosamine, papain(form papaya latex), bromelain(from pineapple stem), degradation reaction.
中文摘要.............................................. I
英文摘要.............................................. II
目錄.................................................. III
圖目錄................................................ V
表目錄................................................ VII
符號說明.............................................. VIII
第一章 緒論........................................... 1
1.1 前言............................................ 1
1.2 幾丁質、幾丁聚醣、幾丁寡醣、葡萄醣胺............ 4
1.3 化學法與酵素法產生幾丁寡醣之比較................ 8
1.4 不同酵素降解幾丁聚醣的比較...................... 10
1.5 Literiture Review of this study................. 12
Chapter 2 Experimental................................ 15
2.1 Materials....................................... 15
2.2 Experiment of apparatus......................... 15
2.3 Analytical apparatus............................ 16
2.4 Experiment flow sheet........................... 17
2.5 Experiment steps................................ 18
2.5-1 Chitosan dissolved in different acid to observe 18
2.5-2 Determination of glucosamine by color change... 18
2.5-3 Degradation of chitosan solution
concentrated process........................... 19
2.5-4 Determination viscosity of Chitosan
in different reaction conditions............... 24
2.5-5 Experiment installations....................... 26
第三章 葡萄醣胺的檢測................................. 27
3.1使用呈色法來檢測葡萄醣胺的定性與定量分析........... 27
3.2使用HPLC來檢測葡萄醣胺............................. 31
3.3使用呈色法與HPLC法來檢測降解產物內
所含葡萄醣胺的定性與定量分析....................... 34
第四章 使用商業化酵素降解己丁聚醣的黏度測試........... 38
4.1己丁聚醣粘度測定之方法............................. 38
4.2己丁聚醣在不同溶劑下的探討......................... 39
4.3鳳梨酵素降解幾丁聚醣在不同的反應條件下的粘度測定... 41
4.3-1不同酵素含量下的黏度測試........................ 41
4.3-2不同轉速下的黏度測試............................ 44
4.3-3不同溫度下的黏度測試............................ 45
4.4木瓜酵素降解幾丁聚醣在不同的反應條件下的粘度....... 46
4.4-1不同酵素含量下的黏度測試........................ 46
4.4-2不同轉速下的黏度測試............................ 48
4.4-3不同溫度下的黏度測試............................ 49
4.5 Bromelain(from pineapple stem)與
Papain(form papaya latex)的純化....................... 50
4.5-1蛋白質酵素純化的動機............................ 50
4.5-2使用SDS-polyacrylamide gel electrophoresis來確定
Bromelain(from pineapple stem)與Papain
(form papaya latex)內含的蛋白質種類.................. 50
4.5-3用DEAE(diethylaminoethyl) column來分離Bromelain
(from pineapple stem)與Papain(form papaya latex)..... 52
第五章 總結與未來展望................................. 54
5.1總結............................................... 54
5.2未來展望........................................... 54
第六章 參考文獻....................................... 55
附錄.................................................. 58
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