研究生: |
陳怡雯 Yi-Wen chen |
---|---|
論文名稱: |
含聚乙二醇雙性高分子之製備與其固定化酵素之研究 Preparation of Poly(ethylene glycol)-Cotaining Amphiphilic Graft Copolymers and Their Effect on the Immobilized enzyme |
指導教授: |
陳崇賢
Chorng-Shyan Chern |
口試委員: |
李振綱
Cheng-Kang Lee 邱信程 Hsin-Cheng Chiu |
學位類別: |
碩士 Master |
系所名稱: |
工程學院 - 化學工程系 Department of Chemical Engineering |
論文出版年: | 2008 |
畢業學年度: | 96 |
語文別: | 中文 |
論文頁數: | 96 |
中文關鍵詞: | 酵素固定化 、雙性高分子 、胰凝乳蛋白酶 、牛血清蛋白 |
外文關鍵詞: | enzyme immobilization, amphiphilic copolymer, α-chymotrypsin, bovine serum albumin |
相關次數: | 點閱:353 下載:4 |
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為了增加酵素的穩定性、方便酵素的回收利用,常會運用酵素固定化的技術改善之。本研究將探討酵素固定化於高分子載體後,造成酵素分子與基質相互作用之催化能力的改變。本實驗以酯交換反應合成一雙性高分子,是以酸鹼敏感性聚丙烯酸 (PAAc) 為主鏈,聚乙二醇(PEG) 作為高分子之側鏈,此雙性高分子則為酵素 (α-chymotrypsin) 固定化之載體。
本論文針對胰凝乳蛋白酵素於固定化高分子前後,以及固定化高分子含量多寡行催化反應之比較,分別以N-Benzoyl-L-Tyrosine ethyl ether (BTEE) 小分子基質及BSA大分子基質,與酵素進行催化水解反應。結果發現當高分子載體中PEG含量增加,則酵素的剩餘活性相對提高,尤其於pH 9下有較高的活性存在;而在與BSA基質作用下,酵素會因固定化高分子中PEG的含量,影響催化作用的分解速率,當在pH 7環境時,於特定PEG含量下其BSA基質被水解速率明顯增加,然而一但PEG含量持續增加反而因立體排斥效應,使酵素不易與大分子基質結合因而影響水解速率。本研究乃經由大、小基質於催化水解反應下之現象差異,了解酵素、載體與基質間各種作用力的消長。
Enzyme immobilization is one of the most important techniques for increasing the stability of enzyme and recycling.
In this study, it is intended to investigate the activity of enzyme after immobilization toward high and low molecular weight substrates. Amphiphilic copolymer comprising poly(acrylic acid)(PAAc) as the backbone and monomethoxy poly(ethylene glycol)(mPEG) as the grafts were synthesized and characterized. The copolymers were used for conjugation with different amounts of α- chymotrypsin as a pH-sensitive immobilized enzyme system. N-Benzoyl-L-Tyrosin ethyl ether was used as a low molecular weight substrate whereas bovine serum albumin was used as a high molecular substrate. Residual activity increased with increasing the mPEG amount and pH. The cleavage rate of bovin serum albumin was accelerated obviously for the experiments carried out in the pH 7 buffer solution. However, the excess amounts of mPEG would induce steric effect, thereby leading to slow cleavage rate.
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