利用微米/奈米尺度類沸石咪唑骨架材料(ZIF-90)包覆大腸桿菌之相關研究

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研究生：	簡琬錡 Wan-Chi Chien
論文名稱：	利用微米/奈米尺度類沸石咪唑骨架材料(ZIF-90)包覆大腸桿菌之相關研究
指導教授：	謝發坤 Fa-Kuen Shieh
口試委員:
學位類別：	碩士 Master
系所名稱：	理學院 - 化學學系 Department of Chemistry
論文出版年：	2020
畢業學年度：	108
語文別：	中文
論文頁數：	69
中文關鍵詞：	金屬有機骨架材料、大腸桿菌、生物複合材料、類沸石咪唑骨架材料-90 、類沸石咪唑骨架材料-8
外文關鍵詞：	Metal-organic Framework, Escherichia coli, Biocomposites, ZIF-90, ZIF-8
相關次數：	點閱：13 下載：0
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金屬有機骨架材料 (MOFs) 因具規則排列的孔洞，而成為目前最新穎的材料之一，主要特點包括高的熱及化學穩定性、尺寸大小可調控性和孔洞之選擇性。已有報導將金屬有機骨架材料塗層於酵母菌細胞表面，藉此抵禦生物裂解酶的攻擊，並使小分子營養乳糖從孔洞擴散進去而提供細胞養分，然而其表面塗層有缺陷，亦可能使抗生素氨苄青黴素等小分子擴散進去而降解細胞。本研究先合成奈米級類沸石咪唑骨架材料-8 (ZIF-8) 塗層於大腸桿菌表面，並證實抗生素確實可能經由缺陷擴散進去細胞，而使細胞裂解死亡。並進一步利用本實驗室於2015年發表之合成方法，利用微米級類沸石咪唑骨架材料-90 (ZIF-90) 封裝大腸桿菌，使其具備更完整的保護以抵禦抗生素，將材料崩解過後，大腸桿菌仍然可以回到原本的生長狀態。並分別量測大腸桿菌和兩種材料ZIF-8/-90之界達電位，藉此證明界達電位之差值確實會影響形成材料之大小，提供未來合成生物複合材料之參考。

Metal Organic Frameworks (MOFs), an emerging class　of porous materials, have become promising materials because of their high thermal and chemical stability, tunable of their pore shape, and size selectivity. There is a report about MOF materials coating on yeast cells to prevent larger cytotoxic molecules, such as lytic enzymes while permit the transport of nutrients necessary for cell viability. However, the defects on MOF-coating may have a chance to enable antibiotic Ampicillin diffuse through and kill the cell. In this study, we demonstrate how to synthesize ZIF-8 coated E. coli and vertify antibiotics could indeed diffuse through and cause E. coli lysis. Herein, we based on our previous report of de novo approach to encapsulate Escherichia coli (E. coli) into ZIF-90 single crystals, E. coli@ZIF-90, as a completely protective structure against harsh conditions. After decomposition of ZIF-90 materials, E. coli immediately regained full functionality. Moreover, we measured the zeta potential of E. coli and ZIF-8/-90 to prove that surface electrostatic potential indeed have a influence on the size of biocomposites.

中文摘要   I
Abstract   II
圖目錄   VIII
表目錄  X
第一章 緒論  1
1-1金屬有機骨架材料   1
1-2類沸石咪唑骨架材料   4
1-3類沸石咪唑骨架材料-8/-90   5
1-4微生物   7
1-5大腸桿菌 ( Escherichia coli )   8
1-6質體 ( Plasmid )   11
1-7研究動機與目的   13
第二章 實驗部分   14
2-1實驗藥品與材料   14
2-2實驗儀器與原理   16
2-2-1 粉末X光繞射儀 (Powder X-Ray Diffraction, PXRD)   17
2-2-2 掃描式電子顯微鏡 (Scanning Electron Microscope, SEM)   18
2-2-3 熱重分析儀 (Thermogravimetric Analyzer, TGA)   19
2-2-4 螢光顯微鏡 (Fluorescence Microscopy)   20
2-2-5 共軛焦雷射掃描顯微鏡 (Confocal Microscopy)   21
2-2-6 聚合酶鏈鎖反應 (Polymerase Chain Reaction, PCR)   22
2-2-7 攜帶型分光光度計 (Ultrospec 10 Cell Density Meter)   24
2-2-8 瓊脂糖凝膠電泳 (Agarose Gel Electrophoresis)  24
2-2-9 界面電位分析儀 (Zeta Potential Analyzer)   25
2-3實驗步驟   26
2-3-1 培養大腸桿菌之相關實驗步驟   26
2-3-2 微米級類沸石咪唑骨架材料-90封裝大腸桿菌 (E. coli@ZIF-90) 之實驗相關步驟   30
2-3-3 奈米級類沸石咪唑骨架材料-8塗層於大腸桿菌之相關實驗步驟   32
2-3-4 奈米級類沸石咪唑骨架材料-90塗層於大腸桿菌之實驗步驟   33
第三章 結果與討論   34
3-1 大腸桿菌之相關鑑定   34
3-1-1 於類沸石咪唑骨架材料-8/-90之活性探討   34
3-1-2 於鹼性環境之活性探討   35
3-1-3 於2-甲基咪唑 ( 2-MI )環境之活性探討   36
3-1-4 於不同培養液之生長狀態探討   37
3-1-5 於不同培養液中抗生素殺菌效果探討   38
3-1-6 於鹽酸與乙二胺四乙酸環境之活性探討   39
3-2微米級類沸石咪唑骨架材料-90封裝大腸桿菌 (E. coli@ZIF-90) 之相關鑑定   41
3-2-1 粉末X光繞射儀鑑定結果   41
3-2-2 掃描式電子顯微鏡鑑定結果   42
3-2-3 熱重分析儀鑑定結果   43
3-2-4 螢光顯微鏡與共軛焦顯微鏡鑑定結果   44
3-2-5 E. coli@ZIF-90於抗生素環境之活性探討   46
3-3奈米級多晶類沸石咪唑骨架材料塗層於大腸桿菌表面之相關鑑定   47
3-3-1 粉末X光繞射儀鑑定結果   47
3-3-2 掃描式電子顯微鏡鑑定結果   48
3-3-3 於抗生素環境之活性探討   49
3-3-4 ZIF-8/-90與大腸桿菌之界達電位探討   50
第四章 結論  51
第五章 參考文獻   52
                                

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