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研究生:謝佑昇
研究生(外文):Hsieh, Yu-Sheng
論文名稱:第一部分:Chitinibacter tainanensis水解幾丁質之機制;第二部分:Chitinibacter tainanensis之抗藥性分析
論文名稱(外文):Part I:The mechanism of chitin degraded by Chitinibacter tainanensis;Part II:Characterization of drug-resistant in Chitinibacter tainanensis
指導教授:施正元施正元引用關係劉昭麟劉昭麟引用關係
指導教授(外文):Shih, Jeng-YwanLiu, Chao-Lin
口試委員:陳錦坤劉昭麟施正元陳錫金
口試委員(外文):Chen, Jeen-KuanLiu, Chao-LinShih, Jeng-YwanChen, Hsi-Jien
口試日期:2012-05-24
學位類別:碩士
校院名稱:明志科技大學
系所名稱:生化工程研究所
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:2012
畢業學年度:100
語文別:中文
論文頁數:152
中文關鍵詞:幾丁質分解酵素幾丁質分解因子覆蓋檢測
外文關鍵詞:chitinolytic enzymesCDFscover assay
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N-乙醯葡萄糖胺為天然多醣類幾丁質水解之最終產物,近年已發現含有許多生理功能,在本土發現能利用菌體將幾丁質水解成N-乙醯葡萄糖胺之菌株Chitinibacter tainanensis,且細菌死亡後仍能分解幾丁質。因N-乙醯葡萄糖胺之產出需倚賴生物體之參與,有別於利用游離酵素作用的觀念,故稱為生物轉化法,其作用機轉係利用幾丁質分解因子將幾丁質水解成N-乙醯葡萄糖胺。
本研究將具有水解幾丁質能力之細菌殘骸利用含0.1% 月桂醯肌胺酸鈉的PBSG萃取幾丁質分解因子,並利用原生膠體觀察發現該因子含有大分子量之聚合物。利用覆蓋式活性測定於原生膠體上,發現於該聚合物相對位置之濾紙有N-乙醯葡萄糖胺酶之活性。由此推知此蛋白質聚合物在水解幾丁質成為N-乙醯葡萄糖胺上扮演著重要的角色。

N-acetylglucosamine (NAG) is the final product from chitin degradation in nature, it is a versatile sugar. Chitinibacter tainanensis identified in Taiwan and its debris can product NAG from chitin degradation. It induced the chitin degrading factors (CDFs) enzymes for chitin degradation was in the surface of the bacterium. The transformation of NAG from chitin was organism-dependent and entitle with biotransformation method.
In the project, CDFs were extracted with 0.1% sodium lauroyl sarcosinate in PBSG from the debris with the enzyme activity. In the cover assay in native gel, the significant band with enzyme activity was observed. It indicated the CDFs existed in the protein complex playing an important role in NAG product from chitin degradation.

明志科技大學碩士學位論文指導教授推薦書 i
明志科技大學碩士學位論文口試委員審定書 ii
明志科技大學學位論文授權書 iii
誌謝 iv
中文摘要 v
英文摘要 vi
目錄 vii
圖目錄 ix
表目錄 x
縮寫對照表 xi
第一章、緒論 1
第二章、文獻回顧 2
2.1 幾丁質及其衍生物簡介 2
2.1.1 幾丁質 2
2.1.2 幾丁聚醣 3
2.2 幾丁質分解酵素 4
2.2.1 簡介 4
2.2.2分類 5
2.2.3 幾丁質分解酵素之作用機制 7
2.2.4 幾丁質分解酵素之活性測定 8
2.3 N-乙醯葡萄糖胺 10
2.4 生物轉化法 11
2.4.1 作用機制 11
2.4.2 CDFs之酵素活性 11
2.5 纖維素 12
2.5.1 纖維素之結構 12
2.5.2 纖維水解酵素 13
2.5.3 纖維水解酶體 13
第三章、實驗規劃 14
3.1 實驗目的 14
3.2 實驗器材 15
3.2.1 儀器 15
3.2.2 藥品 16
3.2.3 試劑 19
3.3 實驗步驟 20
3.3.1 試劑製備 20
3.3.2 酵素粗萃液之製備 20
3.3.3 蛋白質含量之測定 20
3.3.4 酵素活性之測定 21
3.3.5 原生膠體電泳 21
第四章、實驗結果與討論 24
4.1 菌株基本特性 24
4.2 酵素之萃取 24
4.3 幾丁質分解酵素之定量 24
4.4 蛋白質分解酵素之活性測定 25
4.5 覆蓋檢測 25
第五章、結論與未來展望 27
第六章、參考文獻 46

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