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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 羅翊禎 | zh_TW |
| dc.contributor.advisor | Yi-Chen Lo | en |
| dc.contributor.author | 李湘怡 | zh_TW |
| dc.contributor.author | Hsiang-Yi Li | en |
| dc.date.accessioned | 2024-08-16T17:26:44Z | - |
| dc.date.available | 2024-08-17 | - |
| dc.date.copyright | 2024-08-16 | - |
| dc.date.issued | 2024 | - |
| dc.date.submitted | 2024-08-01 | - |
| dc.identifier.citation | 李奕慧. 碳源對 Pantoea vagans M17 表面移行及生物膜形成的影響. 國立臺灣大學生物資源暨農學院食品科技研究所碩士論文. 臺北, 臺灣, 2023.
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Genetic analysis of anti-amoebae and anti-bacterial activities of the type Ⅵ secretion system in Vibrio cholerae. PLoS One. 2011, 6(8), e23876. Zheng, L.; Wang, S.; Ling, M.; Lv, Z.; Lin, S. Salmonella enteritidis Hcp distribute in the cytoplasm and regulate TNF signaling pathway in BHK-21 cells. 3 Biotech. 2020, 10(7), 301. Zhou, G.; Li, L. J.; Shi, Q. S.; Ouyang, Y. S.; Chen, Y. B.; Hu, W. F. Effects of nutritional and environmental conditions on planktonic growth and biofilm formation of Citrobacter werkmanii BF-6. J Microbiol. Biotechnol. 2013, 23(12), 1673-1682. Zhou, G.; Peng, H.; Wang, Y. S.; Huang, X. M.; Xie, X. B.; Shi, Q. S. Complete genome sequence of Citrobacter werkmanii strain BF-6 isolated from industrial putrefaction. BMC Genomics. 2017, 18(1), 765. | - |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/94672 | - |
| dc.description.abstract | 群體感應 (quorum sensing, QS) 允許細菌感知自誘導劑的變化，在細菌間溝通並因應環境變動。自誘導劑的濃度反映族群密度，細菌據以調節基因表現並展現特定集體行為，包含生物膜生成與表面移行。這兩個表型可能促進細菌導致食品腐敗。研究表示 Pantoea spp. 的 QS 系統以 N-酰基-L-高絲氨酸內酯 (N-acyl-L-homoserine lactones, AHLs) 作為自誘導劑。實驗室先前從腐敗香瓜中篩選出 Pantoea vagans M17，具明顯生物膜生成與表面移行能力。為探討 P. vagans M17 的 QS 系統，本研究先利用多重序列比對確認 P. vagans M17 的合成酶 PagI 及受體 PagR 序列，與 P. vagans C9-1 的序列高度相似，且含有保守胺基酸。同時，以 HPLC-MS/MS 鑑定 P. vagans M17 產生的 QS 自誘導劑為 N-butyryl-L-homoserine lactone (BHL)，並添加 BHL 與潛在的 QS 抑制劑 2(5H)-呋喃酮。在 P. vagans M17 的培養液中，BHL 濃度隨著培養時間增加而上升。添加 40 - 140 ppm 的 BHL 雖不影響 P. vagans M17 生長，但可以提升 24 小時的生物膜生成。添加 8 - 40 mM 的範圍之 2(5H)-呋喃酮減緩 P. vagans M17 生長，而當濃度達 32 mM 以上則會抑制生物膜生成。添加 BHL 與 2(5H)-呋喃酮於養分較缺乏之 M9 培養基，可以分別促進與降低 P. vagans M17 表面移行的能力。此外，在第 6 與 24 小時，BHL 與 2(5H)-呋喃酮抑制 pagI 的表現。pagR 與鞭毛蛋白基因 fliC2 的表現情況不受兩者改變。2(5H)-呋喃酮會抑制並延後生物膜調節基因 bssS1 的表達。這些結果顯示，P. vagans M17 的 QS 對生物膜生成及表面移行的調控具有重要性，但其效果會受到環境中營養成分等因素影響。後續研究可以探討 QS 與環境的交互作用對細菌的適應性之貢獻，以及 QS 對生物膜及表面移行的其他重要因子之影響。對 QS 系統更深入的了解，有助於未來尋找合適的方法進行抑制，以減緩細菌導致的食品腐敗並促進食品安全。 | zh_TW |
| dc.description.abstract | Quorum sensing (QS) is used by bacteria to detect changes in the concentration of autoinducers, communicate with each other, and respond to environmental variations. The concentration of autoinducers reflects population density, enabling bacteria to regulate gene expression and exhibit specific collective behaviors including biofilm formation and swarming motility. These two phenotypes may contribute to food spoilage. Studies have shown that the QS system of Pantoea spp. utilizes N-acyl-L-homoserine lactones (AHLs) as autoinducers. Previously, Pantoea vagans M17, isolated from spoiled cantaloupe, was found to have strong biofilm formation and swarming motility. To investigate the QS system of P. vagans M17, we used multiple sequence alignment to confirm that the sequences of AHL synthase PagI and receptor PagR in P. vagans M17 were highly similar to those in P. vagans C9-1 and contained conserved amino acids. Using HPLC-MS/MS, the autoinducer produced by P. vagans M17 was identified as N-butyryl-L-homoserine lactone (BHL). BHL and the potential QS inhibitor 2(5H)-furanone were further added to observe their impact. Concentration of BHL in the culture medium of P. vagans M17 increased with cultivation time. Exogenous addition of 40 to 140 ppm of BHL did not affect the growth of P. vagans M17 but enhanced 24-hour biofilm formation. 2(5H)-furanone suppressed growth within the range of 8 to 40 mM, and inhibited biofilm formation above 32 mM. Addition of BHL and 2(5H)-furanone to the nutrient-deficient M9 medium significantly promoted and diminished the swarming motility of P. vagans M17, respectively. Furthermore, BHL and 2(5H)-furanone reduced the expression of pagI at 6 and 24 hours. The expression of pagR and the flagellin gene fliC2 were not affected by either compound. 2(5H)-furanone decreased and postponed the expression of the biofilm regulatory gene bssS1. These findings suggest that QS is pivotal in governing biofilm formation and swarming motility in P. vagans M17, while other factors such as environmental nutrients may influence the regulation. Future research can investigate the contribution of interactions between QS and the environment to bacterial adaptability. Besides, the effects of QS on other key factors in biofilm formation and swarming motility are worth further study. A comprehensive understanding of QS is expected to aid in devising strategies to mitigate food spoilage caused by bacteria and enhance food safety. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2024-08-16T17:26:44Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2024-08-16T17:26:44Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 論文口試委員會審定書 i
誌謝 ii 中文摘要 iii Abstract iv Graphic abstract vi 目次 vii 圖次 x 表次 xi 附錄次 xii 第一章、前言 1 第二章、文獻回顧 2 第一節、Pantoea spp. 2 第二節、群體感應與其調控之相關表型 4 一、群體感應功能與機制 4 二、群體感應系統之特性分析 6 三、生物膜與表面移行 8 四、第六型分泌系統 11 第三節、群體淬滅 14 第四節、群體感應與生物膜形成及表面移行之關係 16 第三章、研究目的與實驗架構 18 第四章、材料與方法 20 第一節、實驗材料 20 一、實驗菌株 20 二、藥品 20 三、質體、引子與限制酵素 22 四、藥品原液、培養基與溶劑之配製 23 第二節、儀器設備與套裝軟體 28 一、儀器與設備 28 二、套裝軟體 30 第三節、實驗方法 31 一、菌株群體感應蛋白質序列比對 31 二、菌株群體感應基因刪除 31 三、菌株自誘導劑鑑定 34 四、菌株表型分析 36 五、基因表現量分析 38 第五章、實驗結果與討論 44 第一節、P. vagans M17 中群體感應蛋白質之鑑定 44 第二節、P. vagans M17 群體感應基因刪除突變株之建構 52 一、帶有基因上、下游同源片段的重組質體建構 52 二、sacB 的無標記基因刪除 53 第三節、P. vagans M17 中自誘導劑定性與定量 55 一、P. vagans M17 主要產生之自誘導劑為 BHL 55 二、P. vagans M17 分泌之 BHL 含量隨著培養時間增加而上升 65 第四節、群體感應對 P. vagans M17 生長、生物膜生成與表面移行之影響 68 一、群體感應抑制劑減緩 P. vagans M17 之生長 68 二、P. vagans M17 之生物膜生成受到群體感應調控 73 三、群體感應對 P. vagans M17 表面移行之影響 81 第五節、群體感應對 P. vagans M17 基因表現之影響 90 一、引子效率與專一性 90 二、BHL 與 2(5H)-呋喃酮減少 P. vagans M17 中 pagI 之表現 94 三、2(5H)-呋喃酮減少且延後 P. vagans M17 中 bssS1 之表現 98 四、BHL 與 2(5H)-呋喃酮會調節 P. vagans M17 中 hcp1 與 clpV1 之表現 102 第六章、結論與展望 106 第七章、參考文獻 107 第八章、附錄 127 | - |
| dc.language.iso | zh_TW | - |
| dc.subject | 群體感應 | zh_TW |
| dc.subject | 自誘導劑 | zh_TW |
| dc.subject | Pantoea vagans M17 | zh_TW |
| dc.subject | 生物膜 | zh_TW |
| dc.subject | 表面移行 | zh_TW |
| dc.subject | biofilm | en |
| dc.subject | quorum sensing | en |
| dc.subject | autoinducer | en |
| dc.subject | motility | en |
| dc.subject | Pantoea vagans M17 | en |
| dc.title | Pantoea vagans M17 之群體感應調節對運動性和生物膜形成的影響 | zh_TW |
| dc.title | Impact of quorum sensing regulation on the motility and biofilm formation of Pantoea vagans M17 | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 112-2 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 林旭陽;李月嘉;王如邦;林乃君 | zh_TW |
| dc.contributor.oralexamcommittee | Hsu-Yang Lin;Yue-Jia Lee;Reu-Ben Wang;Nai-Chun Lin | en |
| dc.subject.keyword | 群體感應,自誘導劑,Pantoea vagans M17,生物膜,表面移行, | zh_TW |
| dc.subject.keyword | quorum sensing,autoinducer,Pantoea vagans M17,biofilm,motility, | en |
| dc.relation.page | 165 | - |
| dc.identifier.doi | 10.6342/NTU202402835 | - |
| dc.rights.note | 同意授權(全球公開) | - |
| dc.date.accepted | 2024-08-05 | - |
| dc.contributor.author-college | 生物資源暨農學院 | - |
| dc.contributor.author-dept | 食品科技研究所 | - |
| 顯示於系所單位： | 食品科技研究所 | |
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