中文摘要
　　本研究由高效能厭氧產氫污泥篩選出Clostridium butyricum CGS2純菌，初步測試發現其具優異之產氫效能，並以碳源濃度、溫度與pH為主要變因，利用回應曲面實驗設計法（Response surface methodology，簡稱RSM）探討該產氫菌株之最佳產氫條件。首先利用搖瓶批次實驗進行不同碳源種類與其濃度一次因子的測試，實驗中pH值不予以控制，結果顯示以蔗糖20000 mg COD/l為碳源時，有最佳的產氫速率為262.3 ml/h/l，故利用此結果進一步於醱酵槽中針對不同pH值控制與蔗糖濃度的測試，來找尋RSM設計之中心混成實驗的中心點。結果顯示以pH值5.5及蔗糖濃度為20000 mg COD/l時，氫氣產率為最佳(2.25 mol H2/mol sucrose)，且氫氣產量可高達4.9 l，故以此條件為中心點來設計RSM實驗。由RSM法求得氫氣產率之最佳條件為pH 5.2、溫度 35.1oC、蔗糖濃度 22.5 g COD/l，此條件之氫氣濃度為58.5%，其產氫速率為0.54 l/h/l，總產氫量為7.2 l，氫氣產率可達2.91 mol H2/mol sucrose。而以RSM求得產氫速率之最佳條件為pH 5.36、溫度35.1 oC、蔗糖濃度26.1 g COD/l，其氫氣濃度為63.3%，氫氣產率為3.26 mol H2/mol sucrose，總產氫量為10.5 l，產氫速率可達0.50 l/h/l。實驗結果並證實以RSM法所得之最佳化培養條件，確能進一步提升C. butyricum CGS2之產氫效率。

　　隨後利用RSM實驗所得之最佳條件，進行連續醱酵產氫測試。結果發現pH值太低不適合細胞生長，在HRT=12 h時即為成菌體濃度洗出，但若將pH值控制在6.5時則有最佳之產氫效果。因此，便以修正後之醱酵條件進行不同HRT之連續醱酵產氫。結果比照對照組發現當操作於HRT=8 h時，氫氣產率已從0.70 mol H2/mol sucrose提升到5.31 mol H2/mol sucrose，增加了7倍左右，且產氫速率也由0.11 l/h/l提升到0.90 l/h/l，氫氣濃度也大幅度上升為50％。隨著HRT之調降，產氫效率也隨之提升，且當操作在HRT=3 h時，有最佳之產氫速率為1.34 l/h/l，其氫氣產率為 4.40 mol H2/mol sucrose。最後，利用此連續醱酵產氫結果來進行動力學模擬，以不同稀釋速率D(h-1)、細胞濃度X(g/l)、蔗糖濃度S(g/l)、產氣濃度PG,i (mol/l)和液相代謝物濃度PL,i(mg COD/l)為參數來建構此model，而以此model來描述在連續產氫時，各個參數間之相對關係。

Abstract
　In this study, an indigenous Clostridium butyricum CGS2 strain was isolated from highly efficient anaerobic hydrogen-producing sludge. Preliminary tests show that the C. butyricum CGS2 strain exhibited good hydrogen producing activity. Response surface methodology (RSM) was then applied to identify the optimal conditions for hydrogen production of C. butyricum CGS2 using carbon substrate concentration, temperature and pH as the primary operation parameters. First, one-level experiments were done in shake flasks to examine which type and concentration of carbon source was most efficient for hydrogen production. The results show that sucrose at a concentration of 20000 mg COD/l gave the highest hydrogen production rate (vH2) of 262.3 ml/h/l. Based on this result, the effect of pH and sucrose concentration on hydrogen production was further investigated in a fermenter to determine the center point for RSM experimental design. It was found that at a pH of 5.5 and a sucrose concentration of 20000 mg COD/l, the highest hydrogen yield (YH2) of 2.25 mol H2/mol sucrose was obtained, as total hydrogen production was nearly 4.9 l. Hence, the aforementioned conditions was used as the center point for RSM design. Using YH2 as the performance index, the optimum condition predicted from RSM was pH=5.2, temperature=35.1oC, and sucrose concentration=22.5 g COD/l. Under this condition, the hydrogen content was 58.5%, vH2 was 0.54 l/h/l, total hydrogen production was 7.2 l, and YH2 was 2.91 mol H2/mol sucrose. On the other hand, when vH2 was used as the performance index, the optimum condition was pH=5.36, temperature=35.1 oC, and sucrose concentration=26.1 g COD/l. This condition gave a hydrogen content of 63.3%, a YH2 of 3.26 mol H2/mol sucrose, total hydrogen production of 10.5 l, and a vH2 of 0.50 l/h/l. The validity of RSM predictions was confirmed by experimental results, suggesting that using RSM design could attain an optimal culture condition for C. butyricum CGS2 to enhance its hydrogen production performance.
　
　In the next experiments, the optimal culture condition predicted by RSM design was used to perform continuous hydrogen production in a CSTR process. It was found that the pH (5.36) was too low to limit cell grow, resulting in cell wash-out even at a high HRT of 12 h, whereas maximal hydrogen production performance was attained when the pH was controlled at 6.5. Therefore, the culture condition for continuous hydrogen fermentation was modified by using pH 6.5 instead of 5.36. With the modified condition, the reactor was operated at a progressively decreased HRT from 8 h to 2 h. The results show that operation at HRT=8 h allowed a 7 fold increase (from 0.70 to 5.31 mol H2/mol sucrose) in hydrogen yield when compared with control run. The hydrogen production also marked increased from 0.11 l/h/l to 0.90 l/h/l. The hydrogen content increased to 50%. As the HRT decreased, the hydrogen producing efficiency increased. The highest hydrogen production rate (1.34 l/h/l) and yield (4.40 mol H2/mol sucrose) was obtained when the system was operated at HRT=3 h. Finally, the experimental results were subject to numerical simulation with a steady-state kinetic models, and the model appeared to describe the data satisfactorily well.

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