本研究嘗試以biotin選擇性地對Halobacterium salinarium嗜鹽菌紫色細胞膜(purple membrane, PM)胞內外兩側外露之殘基進行修飾鍵結。之後將修飾完成的PM以biotin-strep(avidin)生物親和性吸附方式達成於ITO導電玻璃表面上高度方向性之固定化，製成光電生物感測晶片。利用PM內跨膜之細菌視紫質蛋白(bacteriorhodopsin, BR)受光激發後，會將一質子由胞內側推動至胞外側的光敏感特性，可量測晶片所產生之光電流。首先進行最適化晶片製程的探討，發現以EC端定向之晶片以1.5 mg/mL biotin-PM，經10小時修飾後，可得最佳光電流訊號；CP端定向時則以3 mg/mL之biotin-PM濃度，同樣10小時後最佳。

EC 端定向的晶片在pH=5.75時，有光電訊號極性反轉之現象發生。之後，以QCM感測器分析，QCM晶片頻率的線性下降可證實了多層膜塗覆的成功。最後並藉掃描式電子顯微鏡與原子力顯微鏡觀察其表面形態，確定晶片上具有我們所設計的塗覆。

此研究可應用於製備薄型光感測器或光電開關，並創造未來發展生物性二極體的契機。

The cytoplasmic (CP) and extracellular (EC) sides of purple membrane (PM) from Halobacterium salinarium were each selectively conjugated with biotin for directional immobilization on ITO glass via biotin-streptavidin affinity interactions. The photocurrents of the PM-coated ITO chips, which were produced due to a proton gradient across PM generated by illuminated bacteriorhodopsin, a light-driven proton pump, were studied to investigate the optimal fabrication processes, evaluate the performances of those two differently oriented PM chips, and develop novel photocell systems. The results showed that the best photocurrent responses were achieved when 1.5 mg/mL and 3 mg/mL biotin-conjugated PM were used to coat streptavidin-coated ITO glass for 10 h to prepare the chips with the EC-side and CP-side facing ITO, respectively. In addition, reversion of the photocurrent polarity was observed at pH 5.75 using the chips with the PM EC-side facing ITO. Multilayer fabrication of unidirectionally oriented PM chips was successfully achieved using the developed side-selective biotin conjugation of PM according to the linear frequency decrease responses of layer-by-layer coated QCM chips. Finally, the fabrication of PM on ITO was confirmed by examining the morphology changes of each differently fabricated ITO glass. This study will benefit the future development of PM-film based photosensors, photoswitches, or photodiodes.

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王世育，”細菌視紫質表面修飾及應用於單層膜光電晶片製備之研究”，國立台灣科技大學化學工程研究所碩士論文(2007)

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