摘要：本計畫擬從微觀的奈米製程著手，藉以改善介觀的光電生化感測技術，進而應用到巨觀的生物能源產業。基於上述原由與背景，本計畫的研究主題定為「以普魯士藍奈米晶體發展新型電致色變生物感測器：生質酒精製程中纖維素醣化與發酵反應即時監控應用」，並預計以三年時程（96/8至99/7）完成整體研發工作與系統整合。第一年的研發重點將聚焦於「具電致色變感測技術相容性之普魯士藍奈米晶體修飾透明電極製程開發」，主要的工作內容將包含普魯士藍奈米粒子與奈米管新型製程技術研究，並對奈米結構普魯士藍修飾ITO及FTO電極製程進行最適化，並評估奈米晶體對電致色變性能的提升之具體成效。第二年的研發則以「專一性、高敏度之纖維二糖、葡萄糖及乙醇奈米電致色變酵素電極產製與感測條件決定」為目標，首先我們將以葡萄糖氧化酵素進行奈米普魯士藍晶體酵素電極製程最適化研究，並配合過氧化氫與還原型菸醯胺腺嘌呤二核酸之電致色變感測，決定氧化酵素與脫氫酵素電極進行生化感測時之操作條件，最後利用不同之氧化酵素及脫氫酵素產製專一性、高敏度之纖維二糖、葡萄糖及乙醇奈米電致色變感測電極，並決定同步光電感測之操作條件與完成感測性能綜合比較。第三年，我們計畫將進行「奈米電致色變生化感測裝置組裝與纖維素醣化及發酵程序即時監控應用」，藉由整合可更換奈米電致色變酵素電極之同步光譜電化學微量反應量測槽、具備過濾純化元件及動力來源之流動分析裝置與可穩定輸出訊號之光電偵測模組，組成可攜式之纖維二糖、葡萄糖以及乙醇同步光電生化感測裝置；並以奈米電致色變感測技術進行小型生化反應器中結晶型纖維素醣化與葡萄糖發酵之程序監控與動力學研究，藉以評估不同酵素組成、微生物菌種與操作條件對生質酒精產率的影響。綜言之，本三年期計畫實際對上、中、下游的科學技術進行垂直整合規劃，也藉此機會印證奈米科技的實用性、增加電致色變技術的附加價值並促成傳統生物產業的精密化與升級。
Abstract: This research project plans to utilize the microscopic nanotechnology to improve the mesoscopic optoelectronic biosensing processes and finally applies the outcome to the macroscopic bioenergy industrials. To this end, the project is entitled “Development of novel electrochromic biosensors based on Prussian blue (PB) nanocrystals: Application to real-time monitoring of cellulose saccharification and fermentation in bioethanol processes.” And we will spend three years (2007/8 – 2010/7) to complete the research work and relevant system integration. In the first year, we will focus on “the R&D for electrochromic-sensing-compatible fabrication of PB nanocrystal-modified transparent electrodes.” The details involve the investigation of novel preparative methods for PB nanoparticles and nanotubes, optimization of the preparing parameters for nanostructured PB/ITO and PB/FTO electrodes, and evaluation of the influences of nanocrystals on the enhanced electrochromic performances. In the second year, the main objective of the project will be “the R&D for the fabrication of nanoelectrochromic enzyme electrodes for highly specific and ultrasensitive cellobiose, glucose, and ethanol detection.” At first, we will take advantage of glucose oxidase as a model enzyme to acquire the optimal parameters for preparation of an enzyme/nanocrystal composite electrode. And we will determine the operating conditions for both oxidase-based and dehydrogenase-based sensing electrodes through the detection of H2O2 and NADH, respectively. Then, we will use different enzymes to produce nanoelectrochromic sensing electrodes for both specific and sensitive detection of cellobiose, glucose, and ethanol and will comprehensively compare the performances of the nanoelectrochromic enzyme electrodes. In the third year, our project will aim “to assemble a nanoelectrochromic biochemical sensing device and to apply the instrument to simultaneously monitor the cellulose saccharification and fermentation.” By integrating a spectroelectrochemical cell with a replaceable nano- electrochromic enzyme electrode, a flow analysis chip with filtration and power modules, and an optoelectronic detection system with a steady signal output, we can assemble a portable optoelectronic sensing device for real-time detection of cellobiose, glucose, and ethanol. Then, with the nanoelectrochromic sensing techniques, both saccharification of crystalline cellulose and fermentation of glucose in small-scale bioreactors will be monitored, and the reaction kinetics will be studied, too. Accordingly, the effects of enzyme composition, microbial strain, and operating condition on the yield of bioethanol production will be evaluated. In summary, this three-year project devotes to the vertical integration of upstream, mi
