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標題: 鏈黴菌Streptomyces griseobrunnrus S3產孢作用與抗生活性表現之分子機制
Molecular Mechanism of Sporulation and Antibiotic Production in Streptomyces Griseobrunneus S3
作者: 曾德賜
關鍵字: 生物技術, 植物保護類
摘要: 本研究之主要目的在以應用性及量產技術台灣及美國專利申請中之台灣本土性S.griseobrunneus S3 為模式菌株,釐清與其產孢分化及抗生物質產生有關之信號分子種類以及相關作用的分子基礎,本計畫第一年主要重點在瞭解γ-丁內酯類化合物於供試菌產孢作用中可能之參與作用,試驗中將嘗試由供試菌培養萃取丁內酯類化合物,並以LC/MS/MS, GC/MS/MS 與NMR 等進行結構組成之分析鑑定,另一重點則在於產孢過程同時產生抗生物質種類之分離與鑑定。第二年計畫中將進一步探討產孢過程中已知DasA、與DasR 等調控基因,在本研究液體培養產孢測試系統中可能之參與作用,主要工作重點將由供試菌選殖此二個標的基因,並嘗試瞭解其與γ-丁內酯類信號分子產生與特定醣類分子含量變化間之交互作用關係。計畫中並將一併探討含N-acetylglucosamine供試菌細胞壁分解產物對於產孢分化之影響。第三年計畫中工作重點將深入探討液體醱酵醣類饋料模式與監控參數變化及產孢分化相關性,並對產出孢子與固體培養之孢子進行超顯微結構瞭解,以做為未來生物製劑配方調理改進之依據。本計畫所產出試驗結果將對鏈黴菌繁殖生長過程之產孢分化作用提供具體的生理、生化與分子生物學理論基礎,其對於所發展鏈黴菌生物殺菌劑之產業化發展將甚有裨益。
The main objectives of this investigation is to explore the signal factorsand the molecular basis involved in the sporulation and antibioticproduction of Streptomyces using a Taiwan native S. griseobrunneus S3(Patent Pending: ROC: 92131411, US: 11/708524) as a model strain. Themain effort of first year project will focus on the possible involvement ofγ-butyrolactones (GBLs) as an inducing signal factor of sporulation. Theinvolved GBLs will be extracted and the molecular structure will bedetermined by LC/MS/MS, GC/MS/MS and/or by NMR. Also to beexplored in the first year project is the identification of the antibioticsproduced during the sporulation process. In the second year, continuedeffort will be devoted to further elucidate the participation of knownDasA、DasR genes during the sporulation process. The targeted genes willbe cloned, and their regulatory mechanism in relating to GBLs inductionand cellular carbohydrate content will be examined. The possibleinvolvement of the N-acetylglucosamine contacting bacterial cell walldegrading products in sporulation induction will be also examined as ithas been shown to be critical in inducing antibiotic production. In thethird year project, the ultrastructure of spores produced in the fermentorproduced broth culture will be compared with that by solid culture. Thedifferences observed shall provide valuable information useful for theformulation improvement of biofungicide application. And the possibleconnection of chitinase production and the above studied sporulationsignaling process will be also investigated. The data obtained shall shedthe lights on the myth of the spore differentiation during the reproductivephase which should benefit greatly to the intended development ofStreptomyces biofungicides.
其他識別: NSC99-2313-B005-020
Appears in Collections:動物科學系



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