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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 吳世雄 | |
| dc.contributor.author | Miao-Hsia Lin | en |
| dc.contributor.author | 林妙霞 | zh_TW |
| dc.date.accessioned | 2021-06-13T08:07:48Z | - |
| dc.date.available | 2008-07-28 | |
| dc.date.copyright | 2005-07-28 | |
| dc.date.issued | 2005 | |
| dc.date.submitted | 2005-07-21 | |
| dc.identifier.citation | Aguilar, A., Ingemansson, T., and Magnien, E. (1998) Extremophile microorganisms as cell factories: support from the European Union. Extremophiles 2: 367-373.
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Wood, D.W., Gong, F., Daykin, M.M., Williams, P., and Pierson, L.S., 3rd (1997) N-acyl-homoserine lactone-mediated regulation of phenazine gene expression by Pseudomonas aureofaciens 30-84 in the wheat rhizosphere. J Bacteriol 179: 7663-7670. Wozniak, D.J., Wyckoff, T.J., Starkey, M., Keyser, R., Azadi, P., O'Toole, G.A., and Parsek, M.R. (2003) Alginate is not a significant component of the extracellular polysaccharide matrix of PA14 and PAO1 Pseudomonas aeruginosa biofilms. Proc Natl Acad Sci U S A 100: 7907-7912. 朱聖棻 (2003) Thermus aquaticus NTU103 生物膜形成及質體 pTA103 複製機制之研究. In 慈濟大學醫學研究所 花蓮: 慈濟大學. 李俊蔚 (2003) 灣嗜熱菌株生物模型橙汁蛋白質體學研究專題報告. In 慈濟大學醫學檢驗生物技術系 花蓮: 慈濟大學醫學檢驗生物技術系. 陳懋彥 (2002) 台灣地區嗜熱性細菌之研究. In 國立台灣大學植物學研究所 台北: 國立台灣大學. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/36612 | - |
| dc.description.abstract | 先前研究指出嗜熱菌屬在自然溫泉環境下，並非自由存在於外界，而是以生物膜的型式，緊密附著在週遭的石頭或礦物鹽上，以求取生存。粗萃取生物膜多醣是經由將Thermus aquaticusYT-1此株菌塗在覆蓋有滅菌後的cellophane膜的Thermus固體培養基上，於60 ℃培養48小時後取得。粗萃取的外多醣可經由TSK HW-65F膠體進一步純化，可收集到4個分離的部分，分別稱為231-1、231-2、231-3與231-4，其分子量範圍分佈從10 kDa至500 kDa。藉由酸水解與衍生化反應後可將這四個分離的部分以氣相層析質譜儀與HPAEC-PAD系統測得單醣組成。結果顯示T. aquaticus YT-1株菌的生物膜含有大量的galactose與N-acetyl galactosamine，兩者比例約是1比1。同時也利用氣相層析質譜儀分析主要單醣鍵結確定在231-1樣品中含有：1,3-linked Gal、1,4-linked Gal、1,3-linked GalNAc；在231-2中有：1,4-linked Gal；231-3含有1,4-linked Gal；231-4則有1,4-linked Gal與1,3-linked GalNAc。然而真正確切結構還有待進一步研究探討。
在培養後的T. thermophilus HB-8液體培養基中發現含有促進生物膜生成的因子，即自誘導物。培養後的液體培養基先經過分子量為10 kDa的centricon濾過後，再經由TSK HW-50F的膠體過濾層析系統分離純化，初步可將之分為七個部分進行生物活性分析。結果顯示有兩個分子量較小的部分具有誘導NTU407菌株形成生物膜的能力，之後將會需要更進一步的純化與實驗，進而將分子的結構鑑定出來 | zh_TW |
| dc.description.abstract | In previous studies, bacterial species of genus Thermus living in natural hot springs are firmly attached to rocks and sands by a complex structure called biofilm. The crude biofilm exopolysaccharides (EPS) are obtained from Thermus aquaticusYT-1 plated on sterile cellophane membrane on Thermus agar plate incubation at 60 ℃ for 48 h. The crude EPS passes through TSK HW-65 gel for progressive purification and four major fractions called 231-1, 231-2, 231-3 and 231-4 with molecular weight range from 10 kDa to 500 kDa are collected. After acidic hydrolysis and derivative, the monosaccharide compositions of these four fractions are analyzed by GC-MS and HPAEC-PAD system. The results indicated that the major components of T. aquaticus YT-1 biofilm are galactose and N-acetyl galactosamine with the molar ratio of 1：1. The main glycosidic linkages are also characterized by GC-MS, showing that there are 1,3-linked Gal, 1,4-linked Gal, 1,3-linked GalNAc in 231-1, 1,4-linked Gal in 231-2, 1,4-linked Gal in 231-3 and 1,4-linked Gal, 1,3-linked GalNAc in 231-4. However, the detail structure should be further investigated.
The cultural medium of T. thermophilus HB-8 can induce to form biofilm and show to contain the autoinducer. The cultural medium is incubated overnight and collected by filtering through 10 kDa centricon. Then the filtered cultural medium is subjected to TSK-50F gel filtration chromatography for fractionation. Seven fractions are collected for bioassay. Two fractions with the smaller MW possess bioactivities to promote NTU407 biofilm formation. More studies are under preparation to elucidate their detailed structures. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T08:07:48Z (GMT). No. of bitstreams: 1 ntu-94-R92b46001-1.pdf: 1189854 bytes, checksum: 09fd5b8d97275a8df10bbcfc39fe3124 (MD5) Previous issue date: 2005 | en |
| dc.description.tableofcontents | 中文摘要 I
ABSTRACT II 縮寫對照表 III 第一章、前言 1 一、嗜熱性細菌： 1 二、嗜熱菌的特性及其用途： 2 三、台灣的環境與嗜熱菌之研究： 3 四、生物膜 (BIOFILM) 的介紹： 4 五、嗜熱菌與生物膜： 5 六、生物膜的形成及微生物的組成： 6 七、數量感應系統 (QUORUM-SENSING SYSTEM)與生物膜： 7 八、研究目的： 8 第二章、材料與方法 10 壹、材料： 10 一、 菌種： 10 二、 藥品： 10 三、 儀器： 10 貳、方法： 11 研究一：嗜熱菌外多醣之分析 11 一、實驗流程表： 11 二、嗜熱菌之培養： 11 三、外多醣 (EXOPOLYSACCHARIDE, EPS) 之分離： 12 四、外多醣之純化： 12 五、酚-硫酸法檢測外多醣之含量： 12 六、外多醣醣酸含量檢測： 12 七、氣相層析質譜儀分析外多醣之單醣組成： 13 八、HPAEC-PAD分析外多醣之單醣組成： 13 九、氣相層析質譜儀分析單糖鍵結： 14 十、SMITH DEGRADATION： 15 十一、蛋白質含量檢測： 15 十二、蛋白質切除 (PROTEINASE K DIGESTION)： 16 十三、DIG KIT之醣蛋白檢測： 16 十四、PAS呈色法： 18 十五、銀染： 18 十六、1H-NMR： 18 研究二：自發誘導物之分離鑑定： 20 一、 實驗流程表： 20 二、 製備CONDITION MEDIUM： 20 三、 初步純化： 20 四、 生物膜誘導活性測試： 20 四、 二次純化： 21 第三章、結果 22 研究一：嗜熱菌外多醣之分析 22 一、 酚-硫酸法檢測外多醣之含量： 22 二、 外多醣之純化： 22 三、 外多醣醣酸含量檢測： 22 四、 氣相層析質譜儀分析外多醣之單醣組成： 22 五、 HPAEC-PAD分析外多醣之單醣組成： 22 六、 氣相層析質譜儀分析單糖鍵結： 23 七、 1H-NMR： 23 八、 外多糖粗萃取之蛋白質含量檢測： 24 九、 DIG KIT之醣蛋白檢測與PAS呈色、銀染： 24 研究二：自發誘導物之分離鑑定： 26 一、初步純化： 26 二、生物膜誘導活性測試： 26 第四章、討論與未來探討方向 27 第五章、參考文獻 29 圖表目錄 表一、目前已知之誘導物與生理影響 39 表二、論文中所使用之菌種 42 表三、外多醣組成成分與比例 42 表四、銀染溶液配方 43 圖一、目前已知HOMOSERINE LACTONE 之結構 44 圖二、氣相層析質譜儀分析單糖鍵結反應過程圖 45 圖三、酚-硫酸法檢測外多醣之含量 46 圖四、外多醣之純化 47 圖五、外多醣醣酸含量檢測 48 圖六、HPAEC-PAD分析外多醣之單醣組成 50 圖七、 1H-NMR OF 231-1，300K，400 MHZ 51 圖八、1H-NMR OF 231-2，300K，400 MHZ 52 圖九、1H-NMR OF 231-3, 300K，400 MHZ 53 圖十、1H-NMR OF 231-4，300K，400 MHZ 54 圖十一、1H-NMR OF 231-3，300K，600 MHZ 55 圖十二、1H-NMR OF 231-3，330K，600 MHZ 56 圖十三、2D-TOCSY OF 231-3，330K，600 MHZ 57 圖十四、HMQC OF 231-3，330K，600 MHZ 58 圖十五、HMBC OF 231-3，330K，600 MHZ 59 圖十七、PAS呈色法與銀染之比較 61 圖十八、自誘導物初步純化 62 圖十九、生物膜誘導活性測試 63 | |
| dc.language.iso | zh-TW | |
| dc.subject | 嗜熱菌 | zh_TW |
| dc.subject | 生物膜 | zh_TW |
| dc.subject | Exopolysaccharide | en |
| dc.subject | Biofilm | en |
| dc.title | 嗜熱菌屬生物膜中自誘導物與胞外多醣之性質鑑定 | zh_TW |
| dc.title | Characterization of Autoinducer and Exopolysaccharide in Thermus spp. Biofilms | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 93-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 蔡珊珊,林俊宏,林光慧 | |
| dc.subject.keyword | 嗜熱菌,生物膜, | zh_TW |
| dc.subject.keyword | Biofilm,Exopolysaccharide, | en |
| dc.relation.page | 63 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2005-07-21 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生化科學研究所 | zh_TW |
| 顯示於系所單位： | 生化科學研究所 | |
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