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Microbial Community Structure of Fermentative Hydrogen Production Completely-Mixed Reactor under Various Operating Conditions
|關鍵字:||Fermentative Hydrogen Production;醱酵產氫;CSTR;Clostridium;CSTR;Clostridium||出版社:||環境工程學系||摘要:||
生物產氫中的厭氧醱酵產氫方式因具備不需額外投入能源、代謝率高、所需反應槽體積小並可同時達到廢棄物再利用等特性，因而其產氫效率逐漸受到矚目，其中CSTR厭氧醱酵產氫反應槽已被證實具有良好的產氫效能。本研究利用分子生物技術，PCR - DGGE、cloning以及FISH的方式，探討不同操作條件下CSTR系統中產氫微生物族群的菌相變化。研究結果顯示，當樣本中各菌群比例懸殊時，利用cloning的方式挑選菌群會受到限制；而利用DGGE膠體回收的方式則可順利分析環境中的菌相，由定序比對結果指出在不同的HRT下都有Clostridium的存在，而不同操作條件樣本中還發現K. pneumoniae和K. oxytoca的產氫菌。三種高徑比反應槽在0.5小時之HRT下產氫效果良好，此時槽內污泥形成顆粒狀且利用PCR - DGGE分析當中的菌群變化顯示，不同種類的Clostridium會影響產氫的情形，依照出現菌種的差異，其產氫能力最強者為C. pasteurianum，其次是C. pasteurianum & C. intestinale、C. pasteurianum & C. butyricum，最差者則為 C. butyricum；由FISH結果也發現Clostridium (Chis150) 為優勢菌群，當伴隨有特定比例的Streptococcus (LGC354)時產氫效果最好，而這類細菌多分布於團狀污泥的內部，故推測Streptococcus對顆粒污泥的凝聚扮演重要角色。此外，在高徑比5:1、操作於HRT為0.5小時且蔗糖濃度40000 mg COD/L的反應槽中，C. pasteurianum為主要菌群且其基質轉換率高達3.35 mole H2/mole sucrose。
Dark Fermentation is an important method of biological hydrogen production due to its high conversion efficiency, less energy intensive, less reactor volume and wastes reused. CSTR system in fermentative hydrogen-producing system had been confirmed its fine potential. In this research, we focus on the microbial community structures of CSTR under various operating conditions by using molecular biological techniques, including PCR-DGGE, cloning and FISH. Results indicated that when bacterial species were with a wide gap, screening had been limited. However, elution of specific gene from DGGE gel could correctly analyze the bacterial structure. From sequencing, it had been showed that Clostridium existed when reactor were operated under each different HRT in this research, and other H2-producing microfloras included K. pneumoniae and K. oxytoca had been detected. Reactors of three height-diameter ratio had good hydrogen production at HRT of 0.5 hr, sludge became granular form at this condition. Moreover, results of PCR-DGGE analysis had revealed that efficiency of H2-producing was according to the difference of Clostridium species. The higher hydrogen production potential of Clostridium sp. was C. pasteurianum, and then were C. pasteurianum & C. intestinale, C. pasteurianum & C. butyricum, the lower was C. butyricum. From the results of FISH analysis, it indicated that Clostridium (Chis150) was predominant bacteria, and the H2-producing efficiency was better with a partial Streptococcus which distributed into the inner of the granular sludge; therefore, it was suggested Streptococcus played an important role for coagulation of granular sludge. In addition, when reactor of height-diameter of 5:1 was operated at HRT of 0.5 hr and fed with 40000 mg COD/L sucrose, C. pasteurianum was domain population and the yield of hydrogen was 3.35 mole H2/mole sucrose.
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