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研究生:陳慶瑜
研究生(外文):Chin-Yu Chen
論文名稱:海洋菌株 Alteromonas sp. D5 所產脂肪分解之部分純化及探討
論文名稱(外文):Partial purification and characterization of lipolytic enzymes from a marine bacterium Alteromonas sp. D5
指導教授:謝文陽
口試委員:陳卓昇簡文達李宗徽陳逸民
口試日期:2014-07-03
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:海洋研究所
學門:自然科學學門
學類:海洋科學學類
論文種類:學術論文
論文出版年:2014
畢業學年度:102
語文別:中文
論文頁數:76
中文關鍵詞:Alteromonas海洋細菌脂肪分解&;#37238;熱穩定性鹼穩定性陰離子交換層析螢光活性呈色SDS-PAGE酯化能力耐受清潔劑
外文關鍵詞:Alteromonasmarine bacterialipolytic enzymethermostablealkaline stableanion exchange chromatographyfluorescent zymogramSDS-PAGEesterificationdetergent compatible
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本篇研究從宜蘭大溪漁港中的海水,分離出三株具脂肪分解活性的 Alteromonas species D3、D5 和 D8,其中 Alteromonas D5 所產的脂肪分解&;#37238;具有熱穩定性及鹼穩定性,並經由硫酸銨沉澱及陰離子交換層析將酵素部分純化,最終酵素產量為0.11 %,純化倍率為 15.6。螢光活性呈色顯現出 5 個條帶具有活性,SDS-PAGE 顯示出含量最多的酵素分子量約為 30 kDa,但最高活性的酵素分子量約為4 0 kDa。酵素活性最好的條件為 70℃ 及 pH 8,在 20-80 ℃ 及 pH 6-9 都具有活性;多種金屬離子、抑制劑及界面活性劑的測試用來評估酵素的特性;經由實驗觀察,酵素能催化約 90 % 的棕櫚酸與甲醇反應並轉換成酯類,且酵素與四種市售清潔劑混合處理後能維持 80 % 以上的活性。根據本篇研究,此脂肪分解&;#37238;具有工業應用的潛力。

中文摘要 v
Abstract vi
第一章 前言 1
第一節、細菌所產脂肪分解&;#37238;之分類 1
第二節、脂肪分解&;#37238;的應用 2
第三節、從微生物取得脂肪分解&;#37238;之優點 4
第四節、研究動機 6
第二章 材料和方法 7
第一節、實驗材料與設備 7
第二節、實驗所使用之培養基 9
第三節、細菌樣本 10
第四節、分離、培養與篩選菌株 10
第五節、菌株之分泌脂肪分解&;#37238;檢測 10
第六節、蛋白質定量 11
第七節、脂肪分解&;#37238;活性測試 11
第八節、分離株形態與生理生化測試 12
第九節、分離株之 16S rRNA 序列分析 17
第十節、脂肪分解&;#37238;的純化 19
第十一節、脂肪分解&;#37238;的生化特性分析 22
第十二節、脂肪分解&;#37238;的應用 25
第十三節、統計分析 27
第三章 結果 28
第一節、菌株分離與細菌脂肪分解&;#37238;的活性測試 28
第二節、分離株之脂肪分解&;#37238;活性大小 28
第三節、分離株形態與生理生化特性 29
第四節、分離株之 16S rRNA基因序列分析 30
第五節、分離株之 D5 培養條件測試 30
第六節、脂肪分解&;#37238;的純化 31
第七節、脂肪分解&;#37238;的生化特性分析 32
第八節、脂肪分解&;#37238;的應用 34
第四章 討論 36
第一節、海洋細菌Alteromonas species 分離株的鑑定 36
第二節、脂肪分解&;#37238;的純化和特性探討 37
第三節、脂肪分解&;#37238;的應用 40
第五章 結論 42
參考資料 43


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