細菌視紫質(bacteriorhodopsin，BR)是嗜鹽菌Halobacterium salinarum的一種蛋白質，以紫膜(purple membrane，PM)形式存在。BR結構中含有七個α-helices與兩個β-sheets以穩定的二微結構排列成PM，且受光激發後會進入光循環而以數個光學中間態存在。本研究研究首先將PM以生物親和性吸附固定在載體上，而後藉由石英晶體微天平(quartz crystal microbalance，QCM)和原子力顯微鏡(atomic force microscopy，AFM)分析可觀察到當流動相中加入低濃度0.1 %(w/v)的十二烷基硫酸鈉(sodium dodecyl sulfate，SDS)後會使PM去脂質且使BR結構去折疊化，此時BR仍然被吸附在晶片上。同時發現當受固定化之BR在晶片表面進行去摺疊時，由於液體流過會帶動蛋白質鏈向外延展而影響液體黏度；然而當SDS的濃度高達1 % (w/v)時，一樣會有去脂質及去折疊化影響液體黏度的現象，但同時會破壞BR和晶片間的親和性吸附，最後導致BR隨著流動相流走，無法繼續吸附在晶片上而使晶片完全失去活性。其次，利用自組裝的雷射光/偵測器組合系統可偵測BR主光循環中生命週期最長之M態的衰減情形，以動力學分析發現PM的D96N突變種與膠片包埋形式具有較長的M態生命週長。

Bacteriorhodopsin（BR）is a protein residing in the purple membrane（PM）of Halobacterium salinarum. It is constituted of seven α-helices and two β-sheets, forming a stable two dimensional crystalline structure within PM. Upon photoexcitation, BR undergoes a photocycle containing several optical intermediates. In the first part of this thesis, PM was bioaffinity-immobilized on substrates and then flowed over with 0.1% (w/v) sodium dodecyl sulfate (SDS). The flow-injection quartz crystal microbalance (QCM) analysis revealed PM delipidation and BR unfolding during the 0.1%-SDS flow treatment, with BR still adsorbed on the substrate. The AFM analyses of the SDS-treated PM-coated chips confirmed the observation. Furthermore, the QCM study suggested that the unfolded peptide chain of immobilized BR extended while 0.1% SDS was passing over, changing the apparent viscosity of the flowing fluid. When PM-coated chips were flowed over with 1% SDS, we observed not only PM delipidation, BR unfolding, and changes of the apparent flow-viscosity, but also the detachment of BR from the substrate surface presumably due to the deteriorating effect of 1% SDS on bioaffinity interactions. In the second part, the decay of the M-state, an optical intermediate of BR with the longest life-time, was observed with a laser/photodiode setup. The kinetic analysis revealed that the M-state decay was significantly slowed when BR was either Asp96→Asn mutated or gel-entrapped.

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