臺灣博碩士論文加值系統

先前本實驗室針對Deinococcus radiodurans IR表面two-dimensional crystalline surface layers結構及自組合特性作深入的研究，並開發出簡單可大量快速獲得S-layer fraction (SLF) 的方法。本實驗基於改善微脂體穩定性不足方面的缺點，利用SLF與lipid membrane的interaction，可以簡單且快速在微脂體表面貼覆細菌的表層結晶蛋白，形成SLF coated liposome (SLL)。利用螢光顯微鏡發現SLL具有獨特的發光特性，因此可藉由共軛焦雷射掃描式顯微鏡觀察SLF coated liposome的完整性，以及使用原子力電子顯微鏡觀察奈米級SLL的形態。穩定性實驗方面，SLL與微脂體分別經過酸 (pH 1.5 solution) 處理一段時間，利用庫爾特粒徑分析儀(COULTER Multisizer Ⅱ) 來測量其體積與顆粒數變化，結果顯示SLL具有較佳的耐酸性。另外微脂體面對高張和低張溶液，其體積及顆粒數變化相當大。反觀經過SLF修飾的微脂體具有較佳的穩定性。SLL表面的結晶蛋白構造提供了完整的基質可以連接其他功能性分子，提升藥物輸送的專一性。SLL表面的特殊發光特性亦可當作生物感應器或是診斷用的定量標記(quantitative markers)，增添了微脂粒的應用價值。此外SLL的構造也類似簡單的細菌型態，建立了一個研究細菌membrane protein、ion channel 及演化學方面的model system 。

In the previous study, we have established conditions for isolated subunits of the crystalline cell surface layer (S-layer) protein of Deinococcus radiodurans IR and
efficiently in vitro self-assembly of purified protein. More recently we develop a new method for high-yield preparation of S-layer fraction (SLF). In this work, we are interested in improving liposomes instability. Using the self-assembly principle to attach SLF easily onto liposome surface to form a novel type of liposome (SLL). In basic structural principle SLF coated liposome resemble the cell envelopes. We also observe SLL under confocal laser scanning microscopy and atomic force microscopy. Furthermore, we have investigated the stabilizing effect of SLL towards acid or osmotic challenges. We have demonstrated the SLL are more stable than plain liposome. Such SLL system are close to biomimetic structures that resemble archaeal cells or virus envelopes, may be a model system to study membrane protein, ion channel, and evolution process of bacteria. The high stability of SLL and possibility for immobilizing biologically active molecules on the crystalline array may offer potential in various different liposome applications.

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