細菌視紫質 (bacteriorhodopsin, BR) 存在於紫色細胞膜 (purple membrane, PM) 中，為一具有光電轉換特性之蛋白質，可應用於各種光電裝置。本研究將PM與β-alanine胺基酸、生物辨識分子或無機奈米粒子分別結合製成新型複合材料或晶片，探討其光電與光學性質，並進行各種材料分析。首先將β-alanine與PM溶液混合並製備單晶體，發現PM的混入可使原本β-alanine晶體成為具有光電與非線性二倍頻特性之複合晶體。其次，將抗大腸桿菌 (Escherichia coli) 抗體以avidin-biotin生物親和作用固定化在塗覆有均一方向PM膜的ITO基材上，並進行E. coli捕捉與偵測。發現PM晶片所產生之光電流會因菌被捕捉覆蓋晶片而下降，可應用於菌液濃度定量檢測；相同原理也可應用於一般革蘭氏陰性菌的檢測。再者，將奈米金粒子 (gold nanoparticles, AuNPs)，以生物親和作用或化學鍵結合於塗覆有均一方向PM膜的ITO玻璃上，同樣也發現AuNPs結合濃度增加時會使PM晶片光電流下降，80 nm AuNPs所造成光電流下降效應比10 nm AuNPs顯著；且當10 nm AuNPs塗覆濃度提升至1 µM時，PM晶片可能因AuNPs的SPR效應而造成PM脈衝式光電流的延長，以及PM化學電容效應增加。最後，將綠色量子點 (quantum dots, QDs) 同樣接於塗覆有均一方向PM膜的ITO基材表面上，發現以藍光激發此PM-QDs複合晶片時，可產生連續光電流 (179.6±0.3 nA/cm2)；置換電極為金電極時，則可再提升連續光電流密度至5.7 µA/cm2。進一步，量測BR之M光學中間態的衰減時間常數，發現PM與QDs結合後會縮短，因此推測藍光激發PM-QDs晶片後，QDs先發射出綠螢光而激發BR進入光循環，同時藍光也造成BR M態加速衰退而立即回到基態，如此使BR持續推出質子並累積在PM膜表面而產生連續光電流。利用TEM分析QDs於PM膜上的結合分佈情況，可估算QDs與BR間的螢光共振能量轉移 (Förster resonance energy transfer, FRET) 效率為85 %。此外，使用Maker fringes技術對PM-ITO晶片進行二倍頻量測，可得到PM膜的二階非線性係數值為 χ33(2) = 1.9×10-9 esu且χ31(2) = 1×10-9 esu，證明所塗覆在ITO電極的PM膜具有高度定向性；對於PM-QDs複合晶片，則因QDs的影響而無法測得這些參數。本研究所揭示PM複合材料與晶片之光電與光學特性，可應用於新型生物感測器和生物太陽能電池之開發。

Bacteriorhodopsin (BR) in purple membranes (PM) is a protein with photoelectric conversion property, and can be applied in various photoelectric devices. In this study, β-alanine amino acid, bio-recognition molecules and inorganic nanoparticles were each assembled with PM to form new composite materials or chips, followed by the studies of their photoelectric and optical properties as well as material analysis. First, a single crystal was prepared by mixing β-alanine and PM together and the resultant composite crystal was found to carry photoelectric and nonlinear-optical properties. Secondly, antibodies against Escherichia coli were immobilized on an ITO electrode layered by a uniformly oriented PM layer via avidin-biotin bioaffinity interaction, and then E. coli was captured and detected. The reduction of photocurrent generated by PM was observed due to cell attachment；therefore, the E. coli concentrations were quantified. The same detection principle was further applied to the detection of general gram-negative bacteria. Thirdly, gold nanoparticles (AuNPs) coated on the same unidirectionally immobilized PM layer on ITO also caused PM photocurrent reduction, with the reduction effect of 80 nm AuNPs more significant than that of 10 nm AuNPs. When coated with 1 μM 10 nm AuNPs, the transient photocurrent of the PM chip decreased slower and the chemical capacitance effect of PM increased, possibly due to the SPR effect by AuNPs. Finally, green quantum dots (QDs) were attached to unidirectional PM layer immobilized on ITO. When blue light excited the PM-QDs composite chip, a continuous photocurrent of 179.6±0.3 nA/cm2 was produced. The replacement of the ITO electrode with a gold one further increased the continuous photocurrent density was to 5.7 μA/cm2. The decay time constant of the M state of BR was decreased when QDs were present or attached to. Therefore, it was interpreted that green fluorescence emitted from QDs excited by a blue light drove BR into its photocycle；and meanwhile, the illuminating blue light accelerated the decay of the BR M state, driving BR to quickly return to its ground state. This caused an accumulation of protons on PM surface, resulting in the production of a continuous photocurrent. TEM analysis revealed the arrangement of QDs binding on PM patches and a 67 % efficiency of Förster resonance energy transfer (FRET) between QDs and PM was estimated. Moreover, the second-order nonlinear susceptibilities of the PM-ITO chip were estimated as χ33(2) = 1.9×10-9 esu andχ31(2) = 1×10-9 esu using the Maker fringes method, demonstrating a high uniformity of PM orientation on ITO. The χ33(2) and χ31(2) parameters of the PM-QDs composite chip could not be determined due to the interference of QDs. In conclusion, the revealed photoelectric and optical properties of PM-based composite materials and chips can be applied to the development of new biosensors and biological solar cells.

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