本研究以傳統培養與分子生物學方法，分析纖維素分解過程中微生物社會結構的動態變化且純化纖維素分解菌株。本研究之菌種取自以纖維素為基質的厭氧生物反應槽污泥，最初植種蔗渣堆肥，在60℃利用alpha-cellulose優厚培養數代，優厚培養的菌群能將濾紙分解。16S rRNA基因分子選殖資料庫共有9個OTUs。分解alpha-cellulose的微生物結構，主要由Clostridium屬所構成，其中以Clostridium cluster IV佔40.5%為最多，其次分別為cluster I含29.5%，cluster III則佔24.6％和cluster VII有3.3%的佔有率。Clostridium cluster I 包含許多能產氫的菌株，在厭氧生物反應槽樣本的菌群中，clone SC55-c5與分離的純菌株pure-A1 均和Clostridium thermobutyricum 非常相似。Clone SC55-c4和SC55-c34 與Clostridium cluster IV的菌屬較為相近，其與C. cellulosi 16S rRNA gene的相似度可達98%，分離菌株pure-A4的16S rRNA 基因序列亦與高溫纖維素分解菌株C. cellulosi較為相近。Clostridium cluster VII屬於高溫糖解產氫菌株，clone SC55-c3與分離菌株pure-rca及pure-rcb和Thermoanaerobacterium屬有99%的相似度。另外，本研究亦利用HOPE及T-RFLP偵測纖維素分解過程中菌群結構的動態變化，共測試了cellobiose、avicel cellulose及濾紙，發現在分解初期與分解後期的菌群大不相同，cellobiose分解初期以C. cellulosi為主，分解後期則以Thermoanaerobacterium thermosaccharolyticum為主；avicel cellulose分解初期以C. cellulosi為主，分解中期則以T. thermosaccharolyticum為主，分解後期又以C. cellulosi為主；濾紙分解情況與avicel cellulose相同。

This study used traditional and molecular methods to detect dynamic changes of microbial community structure during cellulose degradation and to isolate cellulose-degrading bacteria. Sample was from anaerobic bioreactor used sugar cane waste compost as seeding and influent with cellulose and the source was sugar cane waste compost. The sludge from reactor was enriched with alpha-cellulose at 60℃. The enriched bacteria can degrade filter paper. In the 16S rRNA gene clone library, there were nine OTUs and alpha-cellulose degrading microbial community belongs to genus of Clostridium. The relative abundance of Clostridium cluster IV, Clostridium cluster I, Clostridium cluster III and Clostridium cluster VII were 40.5%, 29.5%, 24.6% and 3.3%, respectively. Clostridium cluster I includes hydrogen-producing bacteria, clone SC55-c5 and isolated strain pure-A1 are closely to Clostridium thermobutyricum. Clone SC55-c4, clone SC55-c34 and isolated strain pure-A4 are closely to C. cellulosi that belong to Clostridium cluster IV which includes cellulose-degrading bacteria. Clone SC55-c3, isolated strain pure-rca and isolated strain pure-rcb belong to Clostridium cluster VII which includes thermophilic hydrogen-producing bacteria. Besides, we applied both HOPE and T-RFLP to detect dynamic changes of microbial community structure during cellulose degradation. We used three kinds of substrate, including cellobiose, avicel cellulose and filter paper. Microbial populations have changed during cellulose degradation. In the beginning of cellobiose - degrading, the dominant bacterium is C. cellulosi and in the evening of cellobiose - degrading, the dominant bacterium is Thermoanaerobacterium thermosaccharolyticum; in the beginning of avicel cellulose degradation, the dominant bacterium is C. cellulosi. In middle stage of avicel cellulose degradation, the dominant bacterium is T. thermosaccharolyticum and in the evening of avicel cellulose degradation, the dominant bacterium is C. cellulosi. And the changes of microbial populations during filter paper degradation are the same with avicel cellulose degradation.

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黃承泰。本土性梭菌屬纖維素分解菌株之分離及生理特性研究。國家科學委員會計畫期末報告。行政院國家科學委員會，2005。
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