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研究生: 廖清德
Ching-Te Liao
論文名稱: 紫膜生物光電晶片於朝鮮薊抑菌研究之應用
Study of the antibacterial effect of artichoke extract using a purple membrane-based photoelectric biochip
指導教授: 陳秀美
Hsiu-Mei Chen
口試委員: 葉旻鑫
楊顓丞
學位類別: 碩士
Master
系所名稱: 工程學院 - 化學工程系
Department of Chemical Engineering
論文出版年: 2021
畢業學年度: 109
語文別: 中文
論文頁數: 87
中文關鍵詞: 紫膜抑菌朝鮮薊光電晶片
外文關鍵詞: Purple membrane, Antibacterial, artichoke, photoelectric chip
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  • 本論文利用以Halobacterium salinarum紫膜 (purple membrane,PM)為訊號轉換器的生物光電感測器對於朝鮮薊萃取液的抑菌效果進行檢測與探討。由於微生物的遮光性可降低塗覆有PM膜電極晶片的光電流產生強度,因此我們可量化觀察朝鮮薊萃取液添加對微生物的增生或成膜之影響。本論文分為兩個部分,第一部分使用可進行即時檢測之連續注流式PM膜雙層微流道感測器系統,將塗覆有微生物的玻璃置於上層微流道,並連續注入添加不同稀釋比例朝鮮薊萃取液添加量之培養基,同時對置於下層微流道的PM膜電極晶片連續進行光電流量測,如此可即時量化偵測生物膜的生長情形。結果發現,於40 μL/min流速下,流動培養基中添加15倍稀釋之朝鮮薊萃取液時,有最佳的抑菌效果;其中對於Candida albicans、Candida parapsilosis與Escherichia coli的抑制效果最為顯著。第二部分使用可分別檢測真菌、格蘭氏陽性菌與格蘭氏陰性菌的三種PM膜微生物感測晶片,對於靜態培養時添加15倍與25倍兩種不同稀釋比例朝鮮薊萃取液的微生物溶液進行不同培養時間下菌濃度量化檢測。結果同樣顯示培養時添加15倍稀釋之朝鮮薊萃取液有最佳的抑菌效果。抑菌效果於第12小時培養時達顛峰,可維持18小時;但於第24小時即失去效果。本研究顯示朝鮮薊萃取液有明確抑菌效果,且PM膜光電感測器適合應用於微生物生長與培養,以及材料或物質的抑菌性研究。


    We use a biophotoelectric sensor with Halobacterium salinarum (purple membrane ,PM) as the signal transducer to detect and discuss the antibacterial effect of artichoke extract. Since shading the microorganisms can reduce the intensity of the photocurrent generated by the chip, we can quantify the effect of the addition of artichoke extract on the microorganisms. This paper is divided into two parts. The first part uses a continuous injection PM membrane double-layer microfluidic sensor system that can be used for real-time detection. The glass coated with microorganisms is placed in the upper microfluidic channel and continuously injected Different dilution ratios of artichoke extracts are added to the medium, while continuous photoelectric flow measurement is performed on the PM chips placed in the lower microchannel, so that the growth of biofilm can be quantitatively detected in immediate time. The results showed that, at a flow rate of 40 μL/min, when 15 times diluted artichoke extract was added to the flowing medium, the best antibacterial effect was found; Artichoke extract has the most significant inhibitory effect on Candida albicans, Candida parapsilosis and Escherichia coli. The second part uses three types of PM chips that can detect fungi, Gram-positive bacteria and Gram-negative bacteria, for microorganisms with two different dilution ratios of artichoke extract, 15 times and 25 times, added during static culture The solution was subjected to quantitative detection of bacterial concentration under different times. The results also showed that adding 15 times diluted artichoke extract has the best antibacterial effect. The antibacterial effect reached its peak at the 12th hour of culture and could be maintained for 18 hours; however, the effect was lost at the 24th hour. This study shows that the artichoke extract has a clear antibacterial effect, and the PM photoelectric sensor is suitable for microbial growth and cultivation, as well as the antibacterial research of materials or substances.

    中文摘要 II 英文摘要 III 致謝 IV 目錄 V 表目錄 VII 圖目錄 IX 第1章 緒論 16 第2章 文獻回顧 18 2-1 朝鮮薊 ( Artichoke ) 18 2-1-1 朝鮮薊簡介 18 2-1-2 朝鮮薊功效 19 2-1-3 朝鮮薊成分介紹 20 2-2 細菌視紫質 ( bacteriorhodopsin, BR ) 21 2-2-1 Halobacterium salinarum與 BR簡介 21 2-2-2 BR結構 21 2-2-3 BR 光循環路徑 23 2-2-4 BR光電響應 24 2-2-5 PM之單層貼覆及固定化 26 2-2-6 PM晶片的微生物檢測應用 28 第3章 實驗 31 3-1 實驗目的與說明 31 3-2量測 32 3-2-1 Cuvette 系統之D1、D2 微分光電流量測 32 3-2-2 雙層即時監測系統 PM晶片 D1、D2微分光電流訊號量測 33 3-2-3 倒立式螢光顯微鏡 (Olympus IX73) 操作 34 第4章 結果與討論 35 4-1 朝鮮薊萃取液最佳抑菌稀釋比例之探討 35 4-1-1 注流式雙層微流道檢測系統中,培養基流速探討 35 4-1-2 注流式雙層微流道檢測系統中,朝鮮薊萃取液用以抑菌最佳之稀釋比例 37 4-1-3 8種菌株添加朝鮮薊萃取液抑菌之探討 43 4-2 以靜態檢測方式探討朝鮮薊對8株菌之抑菌效果 55 4-3 以靜態檢測方式探討市售漱口水對 S.mutans抑菌效果 69 第5章 結論 74 第6章 參考文獻 76 第7章 附錄 79 7-1 漱口水資料 79 7-1-1 李斯德林漱口水 79 7-1-2 德恩奈漱口水 79 7-1-3 棉花田漱口水 80 7-1-4 高露潔漱口水 80 7-2 螢光圖 80

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