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標題: Development of Artificial Cellulosome for Saccharification(I)
作者: 黃介辰
關鍵字: 應用研究;能源工程, 化學工程類
研究摘要本子計畫將利用纖維素分解酵素複合體之cohesin domains 與dockerin domain之結合關係,配合合成生物學技術以建構出可表現異源性木質纖維素分解酵素之人工纖維素分解酵素複合體,並研發以其複合體為基礎之酵素量產平台。將以耐高溫並在高溫可表現高度活性之C. thermocellum 所產生之cellulosome 為主要架構,利用其anchor protein 與支架蛋白以及其主要木質纖維素分解酵素初步建構人工纖維素分解酵素複合體後再配合其他子計畫所開發之酵素,以酵素工程之方式置入cohesin domains 以期能與支架蛋白結合而形成具異源性基因之人工纖維素複合體,最後再測試整組基因群之酵素量產化條件以建立量產平台。在執行技術上本計畫將採用新興合成生物學技術以突破以往的分子生物技術能轉殖基因之數目所受之局限。宿主將採用 Bacillus subtilis,除了與Clostridium 為親源外,其外泌系統極為發達並可在有氧環境中快速培養成長,非常適合工業化的應用。透過此計畫之執行,除可建構木質纖維素分解酵素表現平台外,更可利用此方法探討何種酵素之組合可達到最佳之分解活性,而這個計畫之目標基因來源不僅僅限定於細菌,亦將由其他子計畫所篩選真菌出有效之纖維素分解脢,再藉由此方法討論不同宿主來源酵素活性間的組合,以增加各種料原之多元性降低料源成本。

bstractThe utility of synthetic biology in this project,binding cohesin domain withdockerin domain, can construct a artificial lignocellulose hydrolitic complex expressheterologous genes on the designer cellulosome as a enzyme production system.The cellulosome of C. thermocellum is a thermostable enzyme and show highefficiency in he high temperature. The complex is including the anchor protein、thescaffolding and cellulolytic enzymes on it, and other heterologous enzyme developedfrom sub-projects will be added in cohesion domains to develop a cross-speciesdesigner cellulosome enzyme complex The gene cluster will be test to establish aenzyme mass production platform. It’s capable to improve the number of transgenicgenes by using synthetic biology. Bacillus and Clostridium are closed bacterium. Itcan growth fast at enzyme production in industry applications. Except the enzymesystem, the best enzyme combination of saccharification will be developed. Thesouces of lignocellulose hydrolitic genes in this project not only focus on bacteria butalso fungus, and the enzyme combinations between different host will be discussedwhether microbes can express high level lignocellulose hydrolitic enzyme moreefficiently ,and increasing sources of subtracts will cause cost down .
其他識別: NSC99-ET-E005-002-ET
Appears in Collections:生命科學系所

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