本研究開發一套新穎的光纖光生物反應器進行光醱酵產氫，由於傳統的光纖材料屬於端點發光，本研究利用表面改質製備側光光纖，並將側光光纖插入光生物反應器作為內部光源，以促進光源效率與分佈。本研究首先探討不同光源對光合產氫效能的影響，結果顯示，在單一光源的測試，使用鹵素燈當作光源可以得到較佳的光合產氫效能，其總產氫速率以及氫氣產率分別為8.68 ml/l/h以及1.09 mol H2/mol HAc。此外，藉由適當的光源組合可提升光醱酵產氫效率，結合雙光源之結果顯示，結合內部光源 (OF-MH)以及外部光源(HL)可將總產氫速率以及氫氣產率分別提升至 17.24 ml/l/h以及2.22 mol H2/mol HAc。有鑑於此，本研究更致力於發展更有效率且更經濟之光醱酵產氫技術，以降低其商業化應用之門檻。
為了瞭解不同操作模式對產氫效能的影響，本研究以批次、連續攪拌式以及fill and draw (F/D)之進料策略探討各操作策略對光合菌產氫效能的影響。在相同的培養條件下(鹵素燈為光源)，使用F/D當作操作策略，在連續10天的操作期間，確實能夠得到較高的光合產氫效率，其產氫速率達15.5 ml/h/l。本研究並以不同濃度(1000-3000 mg COD/L)的乙酸(碳源)以及不同濃度(100-800 mg/L)的肤胺酸(氮源)探討基質組成對該光合菌產氫之影響，結果顯示在最佳的乙酸 (2000 mg COD/l)以及肤胺酸 (400 mg/l)濃度下，其最大產氫速率以及氫氣產率將分別為 20.9 ml/l/h以及 2.47 mol H2/mol HAc。將最佳光源配置應用於F/D之操作策略，其總產氫速率以及氫氣產率大幅提升至 38.2 ml/l/h和 3.15 mol H2/mol HAc。因此，證明本研究所開發之光纖生物反應器與 F/D之操作策略確實能夠有效提升Rhodopseudomonas palutris WP3-5之光合產氫效能。
此外，本研究亦探討固體載體添加對Rhodopseudomonas palutris WP3-5菌株光合產氫效能之影響。添加不同的載體，如活性炭(AC)、矽膠 (SG)以及黏土(clay)於醱酵培養基中可促進菌體濃度以及光合產氫效能，尤其是添加2%的clay能有效提升光合產氫速率(67.2%)以及氫氣產率(37.2%)。接著，本研究以不同濃度(1,000-3,000 mg COD/l)的乙酸，探討使用clay當作載體對光合菌產氫之影響。結果顯示使用1,000 mg COD/l的乙酸當作碳源，總產氫速率以及氫氣產率可分別提高至28.5 ml/l/h及74.3% (2.97 mol H2/mol HAc)。最後，本研究企圖將新穎的光纖發光技術結合clay之添加以及最佳乙酸初始濃度等以促進光合產氫效能，此最佳組合可分別將總產氫速率以及氫氣產率再提升至43.8 ml/l/h 與 3.63 mol H2/mol HAc，此產氫效能明顯高於相關文獻之效能，因此，更說明本研究所開發之光纖光生物反應器確實具有提升光合產氫效能之功效。
雖然改良光反應器系統與操作策略可有效提升光合產氫效能，然而使用人工光源(鎢絲燈、鹵素燈以及副金屬燈)具有高操作成本(燈源電費)以及高建構成本(燈源成本)的缺點。因此，改良光源供應的方式將是發展光醱酵產氫程序的關鍵技術。有鑑於此，本研究以太陽能取代人工光源來激發光纖，並以太陽能光纖光生物反應器來降低大量的電能消耗。結果顯示，利用太陽能激發光纖之發光系統，其累積產氫量以及氫氣產率分別比使用鎢絲燈提高近138% 以及 136%。然而，利用太陽能激發光纖時有光強度不穩定的現象，因太陽能之光照強度會隨著天氣、季節、設置位置、太陽能光譜以及操作時間而改變。因此，本研究開發一套光感測系統去監控反應器所接受之光照強度，當太陽光光纖光生物反應器之內部光照強度低於30 W/m2時，系統會自行開啟光生物反應器外部所架設之鎢絲燈源，以達到均勻且穩定的光能供應。結合光感測器以及太陽能光纖光生物反應器，其累積產氫量以及氫氣產率分別比單獨使用太陽能光纖光生物反應器高出26.8%與26.7%，且電能之消耗更節省。
由於乙酸以及丁酸為暗醱酵產氫之主要液相代謝產物，因此，此兩種碳源的濃度比例被視為光合菌產氫以及菌體生長之重要影響因子。本研究利用回應曲面實驗設計法探討乙酸以及丁酸之最佳組成，並且以氫氣產率 (YH2)、最大氫氣產生速率 (Rmax) 以及總產氫速率(Roverall)為功能指標，來評估光合產氫之效能。由於發現不同功能指標得到不同的乙酸與丁酸最適化組成，為綜合考量三個功能指標(Rmax、Hmax以及 YH2)，本研究開發overlay contour plots 之實驗設計法，已獲得同時滿足Rmax、Hmax與YH2時之最適化碳源比例。
最後，本研究以實際暗醱酵程序所產生的廢水為基質來進行光醱酵產氫程序，藉由兩系統的結合，來提高氫氣產率並降低放流水COD含量。本研究以20000 mg COD/l的蔗糖為碳源，使用由高效能厭氧產氫污泥槽中篩選到的 Clostridium pasteurianum CH4純菌株進行暗醱酵產氫，可得氫氣產率為2.5 mol H2/mol sucrose。接著，將暗醱酵殘留液適當稀釋後(丁酸，乙酸與氨氮濃度分別為2900 mg COD/l、900 mg COD/l以及21 mg/l)作為基質，利用 Rhodopseudomonas palustris WP 3-5進行光醱酵氫氣。結合暗醱酵與光醱酵之產氫程序可將氫氣總產率由原先的2.5 mol H2/mol sucrose提升至7.5 mol H2/mol sucrose，若再結合測光光纖發光系統，其氫氣產率可進一步提高至9.76 mol H2/mol sucrose。因此，結合暗醱酵以及光醱酵之串聯產氫系統，確能有效改進產氫效能並分解暗醱酵產氫廢水所殘留之COD，以避免衍生之環境污染問題。

In this work, a novel optical-fiber-based photobioreactor was utilized to produce H2 with an indigenous photosynthetic bacterium Rhodopseudomonas palustris WP3-5. Plastic cladding of conventional optical fibers was removed to obtain side-light optical fibers (SLOF), which was inserted into photobioreactors as the internal light source. The effect of light sources on the phototrophic hydrogen production was examined. The results show that among the single light source tested, illumination with halogen filament lamp gave a higher overall H2 production rate of 8.68 ml/l/h and H2 yield of 1.09 mol H2/mol HAc. Binary combination of external and internal light sources appeared to give higher photo-H2 production performance, especially when optical fibers (OF-MH) and external lamps (HL) were combined. This motivated us to develop viable fermentation technology to produce hydrogen in a more efficient and cost-effective way.
To determine which operation mode was preferable for the photo-H2 production, batch, CSTR and fill & draw (F/D) operations were conducted. The results suggest that under similar culture conditions, the F/D operation seemed to attain better overall H2 production rate (15.5 ml/l/h) than the other operation modes examined. For medium improvement, the PBR was conducted under different concentrations of carbon source (HAc) and nitrogen source. The results show that the optimal HAc (2000 mg COD/l) and glutamic acid (400 mg/l) concentration led to a vH2 and YH2 of 20.9 ml/h/l and 2.47 mol H2/mol HAc, respectively. Combined with F/D operation strategy and optimal light source arrangement (i.e., the OF/HL/TL system), the overall H2 production and H2 yield was further enhanced to as high as 38.2 ml/h/l and 3.15 mol H2/mol HAc, respectively. These results demonstrated that using optical fiber-illuminating photobioreactor was excellent in promoting H2-producing performance from fill-and-draw cultures of R. palustris WP3-5.
Meanwhile, it was found that photo-H2 production could be improved by adding solid carriers in the fermentation broth containing cells of R. palustris WP3-5. Addition of a small quantity (2%) of clay into fermentation broth significantly promoted H2 production, resulting in 67.2% and 37.2% increases in H2 production rate (vH2) and H2 yield (YH2), respectively. Moreover, the preferable carbon and nitrogen sources was determined, as using a optimal initial HAc concentration of 1000 mg COD/l and glutamate concentration of 400 mg/l led to a vH2 and YH2 value of 28.5 ml/h/l and 2.97 mol H2/mol HAc, respectively. Finally, optical-fiber illuminating system was designed to facilitate the efficiency of the photobioreactor. Combination of internal optical-fiber illumination system, clay addition, and optimal HAc concentration further elevated the vH2 and YH2 to a maximum level of 43.8 ml/h/l and 3.63 mol H2/mol HAc, respectively. These values are considerably higher than most reported results from relevant studies.
A novel solar-energy-excited optical fiber photobioreactor were designed and constructed to promote the phototrphic H2 production of R. palustris WP3-5. The photobioreactor was illuminated by combinative light sources including optical fiber excited by solar energy (internal light source) as well as external irradiation of tungsten filament lamp. The H2 production performance of the photobioreactor was assessed by using different light source arrangement for the day/night illumination. First, a solar-energy-excited optical fiber photobioracctor with a working volume of 50 ml was applied to investigate the effect on the cell growth and photo-H2 production. Second, the reactor was scaled up and different light source arrangement was employed. Finally, an innovative light sensor (light dependent resistor, LDS) was utilized to monitor solar light intensity. In this way, continuous and stable light supply to the photo-bioreactor was achieved by supplementing external light source when the light density from sunlight was low. This design appeared to facilitate the photo-H2 producing stability and productivity.
In the combinative process of dark and photo fermentation, the predominant soluble metabolites (i.e., HAc and HBu) from dark fermentation was used as the substrate for photo-hydrogen production with R. palustris WP3-5. The best composition of HAc and HBu for H2 produciton was determined by using response surface methodology (RSM). In addition, a new overlay contour plots method was used to determine optimization of the HAc and HBu concentration by simultaneously considering all the three performance indexes (i.e., Rmax, Hmax and YH2). Using this method, the culture attained a Rmax of 39.5 ml/h, a Hmax of 2738 ml and a YH2 of 51.6 %.
Dark and photo-H2 production systems have been combined in series to produce H2, achieving a higher yield and a lower chemical oxygen demand (COD) in the effluent. In this study, a pure strain of Clostridium pasteurianum CH4 isolated from efficient anaerobic H2-producing sludge was used for dark fermentation from sucrose medium. C. pasteurianum CH4 gave a H2 yield of 2.5 mol H2/mol sucrose in a batch culture. The dark fermentation broth was further utilized by R. palustris WP 3-5 to produce H2. The total H2 yield increased from 2.50 mol H2/mol sucrose in dark-fermentation to 7.50 mol H2/mol sucrose by using the two-step process. Moreover, combination of side-light optical-fiber illumination system further enhanced the total H2 yield to 9.76 mol H2/mol sucrose. Therefore, combining dark- and photo-fermentation could effectively increase H2 production yield and reduce the COD content derived from dark fermentation effluents.

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