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研究生:劉宛佐
研究生(外文):Wan-Tso Liu
論文名稱:環境中分離紫色非硫光合菌及其可能應用之探討
論文名稱(外文):Isolation of environmental photosynthetic purple non-sulfur bacteria and their potential applications
指導教授:簡志青
指導教授(外文):Chih-Ching Chien
學位類別:碩士
校院名稱:元智大學
系所名稱:生物科技與工程研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2011
畢業學年度:99
語文別:中文
論文頁數:46
中文關鍵詞:紫色非硫紫色非硫
外文關鍵詞:purple non-sulfur bacteria、Carotenoid、Coenzyme Q10
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紫色非硫光合菌 (purple non-sulfur bacteria) 在不同環境中可實行不同代謝方式,例如: 光合自營、光合異營、化學自營、化學異營。由於這類的細菌有其自有的生理特性,在加上光合菌本身廣泛之應用於環保、水產養殖、農業種植、畜牧、醫藥等行業, 最近幾年已受到許多研究者的重視。紫色非硫光合菌廣泛的生長在自然界中,不需要太多的養分,只需一些有機物質以及有光照之地便能生長。在本研究中,我們從北台灣附近水池、稻田、水溝等地的土壤或沉積,利用厭氧光照方式培養,分離光合菌株,利用PCR技術擴增分離菌株之16s rDNA,經由定序分析結果,篩選所要進行探討知紫色非硫光合菌(Rhodobacter sp.)。並評估這些本土分離之紫色非硫光合菌株在應用方面之研究,包括Coenzyme Q10含量的分析、細菌色素含量及種類如類胡蘿蔔素等。

Purple non-sulfur bacteria (PNSB) possess diverse metabolic pathway to survive in different environments, including photoautotrophic, photoheterotrophic, chemoautotrophic as well as chemoheterotrophic metabolisms. PNSB have been widely used in the fields of environmental protection, agricultural production, aquaculture and medical application, etc. The unique physiological characteristics of PNSB have also attracted the attention of researchers to investigate the possible application of these microorganisms. In this study, we have isolated and identified several strains of photosynthetic bacteria from sediments or soil from different environment in northern Taiwan, including pond water, rice field and channel. Enrichment technology was employed for isolation of PNSB with anaerobic incubation under light condition. The isolates were identified by 16S rRNA gene sequence analysis and several strains of Rhodobacter sp. were chosen for further evaluation for their applications such as production of coenzyme Q10 and bacterial pigments.

中文摘要 i
Abstract ii
誌謝 iii
目錄 iv
表目錄 vi
圖目錄 vii
第一章 緒論 1
1-1 前言 1
1-2 動機 2
第二章 文獻回顧 3
2-1光合菌簡介 3
2-2 種類 4
2-3 Coenzyme Q10 5
2-3-1 CoQ10 的簡介 5
2-3-2 CoQ10 的應用 6
2-3 生產CoQ10 6
2-3-1 生產CoQ10的方式 6
2-3-2 生產CoQ10的菌種 7
2-4 類胡蘿蔔素 7
第三章 材料與方法 9
3-1 論文架構 9
3-2 實驗材料 10
3-2-1菌株 10
3-2-1-1環境液體樣本來源 10
3-2-1-2環境泥土樣本來源 10
3-2-2藥品及廠牌 10
3-2-3實驗儀器 12
3-2-7培養基 13
3-3 實驗方法 18
3-3-1 菌種採樣 18
3-3-2 菌種培養 18
3-3-3 菌種分離 18
3-3-4 菌種保存 19
3-3-5 微生物生長之測定 19
3-3-6 染色體DNA純化(Purification of chromosomal DNA) 19
3-3-7 聚合酶鏈鎖反應 ( Polymerase chain reaction, PCR ) 20
3-3-8瓊脂膠體電泳分析(Agarose Gel Electrophoresis) 20
3-3-9瓊脂膠體DNA片段純化(DNA fragment gel extraction) 21
3-3-10 接合反應 (Ligation reaction) 21
3-3-11 轉形作用 (Transformation) 21
3-3-12 藍白菌落挑選 (blue/white screening) 22
3-3-13 質體DNA純化 (Plasmid DNA purification) 22
3-3-14 限制酶切割(Restriction enzyme digest) 22
3-3-15 序列比對 23
3-4 分析方法 23
3-4-1 CoQ10 的檢量線建立 23
3-4-2 CoQ10 檢量線 23
3-4-3 HPLC (High-performance Liquid Chromatography)分析 24
3-4-4 CoQ10 含量分析 24
3-4-5 光合菌色素分析 25
第四章 結果與討論 26
3-1菌株之選殖 26
3-2菌株培養 26
3-3菌株生長情況 27
3-4 菌種鑑定 28
3-5 自行篩選之菌株結果 29
3-6 光合菌色素分析 30
3-7自行篩選之菌株之CoQ10含量分析 31
3-8 不同培養基下與食工所菌株之比較 32
3-9菌液體積下與食工所菌株之比較 32
3-10 不同溫度下 Rhodospitllum rubrum 之比較 33
3-11 不同溫度下 Rhodopseudomonas palustris 之比較 34
3-11利用不同溫度下使用 N.S medium 之菌株生長情形比較 34
3-12不同培養基下菌株生長情形之比較 35
第四章 結論 38
第五章 參考文獻 39
第六章 附錄 44



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