臺灣博碩士論文加值系統

氣候環境溫室效應問題的惡化，與石化燃料的耗竭嚴重影響了人類的生活，故發展其他替代能源已成為現今重要課題。氫氣是一種乾淨且富有高能量(122 kJ/g)的永續能源，目前被視為未來主要替代能源之ㄧ，如能有效的大量生產氫氣，將會對環境有正面的幫助，現階段氫氣生產的方法為石化燃料產生氫、水電解產生氫氣與生物方法產生氫氣。
生物產氫優勢為可利用有機廢棄物、廢水做為碳源，在獲得氫氣的同時淨化了水質且達到能源回收再利用之目的，故本研究為探討利用生物方法產生氫氣。現今具有產氫能力的微生物有綠藻、藍綠細菌、紫色不含硫光合細菌與厭氧菌，由於藍綠菌、紫色不含硫光合細菌與厭氧菌各有不同的產氫條件，能夠有效結合三種微生物達到提升生物產氫量為一重要的研究課題，本研究第一部分是以廚餘作為厭氧產氫基質，利用厭氧暗醱酵系統產生氫氣，再將厭氧暗醱酵含有機酸之出流水提供紫色不含硫光合細菌產氫；第二部分為探討紫色不含硫光合細菌與藍綠細菌共培養產氫之可行性。
實驗先由台中某生活污水廠污泥經由熱篩程序，分離出厭氧產氫菌，並由BMP培養基增殖，作為厭氧暗醱酵槽之植種，再以廚餘進行厭氧醱酵菌半連續式產氫試驗並將其液態產物作為光合細菌連續流產氫試驗，進行兩階段生物產氫，結果顯示利用廚餘作為厭氧暗醱酵之基質能夠產氫，最高累積248ml氫氣，且富含有機酸之出流水氨氮濃度大約為5mg/L，低於紫色光合作用細菌Rhodopseudomonas plaustris WP3-5產氫時氨氮抑制濃度17mg/L，故此厭氧暗醱酵出流水適合後續光合連續流產氫試驗。利用厭氧暗醱酵出流水做為紫色不含硫光合菌產氫基質，最高氫氣產量為988.9 ml-H2/day。
第二部份以紫色光合作用細菌與藍綠菌共培養，其中紫色光合作用細菌以有機酸作為碳源產生氫氣，藍綠菌以異營方式可利用果糖作為碳源來產生氫氣，經由實驗結果得知將紫色光合作用細菌與藍綠菌共培養，可產生氫氣且藍綠菌以果糖產氫會有乙酸及其他代謝物產生，可持續提供紫色光合作用細菌利用而提高氫氣累積量。本試驗設計為將單獨培養紫色不含硫光合細菌、單獨培養藍綠細菌及共培養紫色不含硫光合細菌與藍綠細菌在同一系統條件下進行產氫批次試驗。試驗結果，單獨培養之紫色不含硫光合細菌與單獨培養之藍綠細菌個自累積產氫量相加後之總累計產氫量為56.77 ml，紫色不含硫光合細菌WP3-5與藍綠細菌CH3混合比為1:1時累積產氫量為83.04 ml，而在共培養中，當紫色不含硫光合細菌WP3-5與藍綠細菌CH3混合比為2:1時，為產氫最佳混合比，其最大氫氣累積量為140.83 ml，為兩單獨培養之總累積產氫量之2.48倍，顯示將紫色不含硫光合細菌與藍綠細菌共培養於含有機酸及果糖的培養基中，可以得到比兩者單獨培養時更多的產氫量。

Nowadays, the aggravation of greenhouse effect by combustion of non-renewable sources of energy such as fossil fuel, coal, oil, and nature gas has become a global issue. However, hydrogen can be an alternative and sustainable energy source due to the properties of clean and high energy yield（122 kJ/g）, which is an environmentally friendly technology in the future. Biohydrogen, comparing with the other technologies of hydrogen production, has the advantages of organic waste recycling and cost- effectiveness. Therefore, the aim of this study focuses on the combination of three different types of microorganisms and trying to enhance the efficiency of hydrogen production. This study was divided into two parts. Firstly, after anaerobic fermentation of leftovers, the possibility of using effluent as substrate for phototrophic hydrogen production was evaluated. Secondly, the feasibility of biohydrogen production from co-culture of purple nonsulfur bacterium, Rhodopseudomonas palustris WP3-5, and heterocyst-forming filamentous cyanobacteria, Anabaena sp. CH1, was estimated.
In the first part, the inoculum of anaerobic fermentation was isolated from the sludge from wastewater treatment plant via heat treatment and BMP medium enrichment. The results show that the maximum cumulative volume of biohydrogen from semi-continuous anaerobic fermentation system is 248 ml, which is utilizing the leftovers as substrates. The results also indicate that the effluent contains large amount of volatile fatty acid but little amount of ammonium（5 mg/L）. Because the ammonium concentration in effluent is lower than the inhibition threshold（17 mg/L）of Rhodopseudomonas plaustris WP3-5 during biohydrogen production, phototrophic biohydrogen system could be successfully compatible with anaerobic fermentation system as two-stage biohydrogen production system.
In the second part, the results from co-cultured system show that the maximum cumulative volume of biohydrogen reaches 140.83 ml in the WP3-5/CH1 mixed ratio of 2/1, which increases 2.48 fold comparing with the sum of cumulative volume（56.77 ml）from individual biohydrogen systems. Acetate, the metabolite of fructose via heterotrophic metabolism of Anabaena sp. CH1, could be taken by Rhodopseudomonas plaustris WP3-5 as energy source to produce biohydrogen. Therefore, the results from this study indicate that the co-culture biohydrogen system combining those two phototrophic bacteria is exactly feasible and the efficiency of biohydrogen production by co-cultured system is better than that of the single culture system.

目 錄
中文摘要 I
英文摘要 Ⅲ
目錄 Ⅴ
表目錄 Ⅸ
圖目錄 Ⅹ
第一章 前言 1
第二章 文獻回顧 3
2-1 全球環境與能源危機 3
2-2 可再生能源 4
2-3 氫能源 5
2-4 氫能源的利用 5
2-5 微生物產氫 6
2-5-1厭氧醱酵產氫 8
2-5-1-1厭氧醱酵菌 8
2-5-1-2厭氧醱酵產氫代謝機制 10
2-5-1-3影響厭氧醱酵產氫之因素 13
2-5-2紫色不含硫光合細菌 15
2-5-2-1紫色不含硫光合細菌光合作用與產氫機制 16
2-5-2-2影響紫色不含硫光合細菌產氫之因素 18
2-5-3藍綠細菌 21
2-5-3-1藍綠細菌光合作用與產氫機制 22
2-5-3-2影響藍綠細菌光合產氫之因素 25
2-6結合不同產氫微生物之研究 27
2-7生物產氫之基質 30
第三章 材料與方法 33
3-1實驗架構 33
3-2菌種來源 34
3-2-1厭氧醱酵細菌來源 34
3-2-2紫色不含硫光合細菌來源 34
3-2-3藍綠細菌來源 35
3-3菌種保存與培養 35
3-3-1厭氧醱酵細菌 35
3-3-2紫色不含硫光合細菌 37
3-3-3藍綠細菌 39
3-3-4藍綠細菌與紫色不含硫光合細菌共培養 40
3-4實驗方法 41
3-4-1半連續式厭氧暗醱酵產氫試驗 42
3-4-2 連續式光合產氫試驗 44
3-4-3紫色不含硫光合細菌與藍綠細菌於共培養系統產氫可行性試驗 45
3-4-3-1單獨培養紫色不含硫光合細菌與藍綠細菌產氫批次試驗 45
3-4-3-2共培養紫色不含硫光合細菌與藍綠細菌產氫批次試驗 47
3-5分析設備與方法 49
3-5-1 O.D.值（Optical density）與細胞乾重 49
3-5-2 pH 50
3-5-3 sCOD 50
3-5-4 NH4+濃度 50
3-5-5可溶性碳水化合物濃度 51
3-5-6果糖濃度 52
3-5-7有機酸濃度 52
3-5-8氣體組成 53
3-5-9血球計數器 54
3-5實驗用水與鹼洗液 55
第四章 結果與討論 56
4-1細胞密度與細胞乾重 56
4-1-1菌株Rhodopseudomonas palustris WP3-5 細胞密度（O.D.660）與細胞乾重 56
4-1-2菌株Anabaena sp. CH3 細胞密度（O.D.900）與細胞乾重 57
4-2厭氧醱酵產氫系統 58
4-2-1菌種篩選試驗 58
4-2-2廚餘厭氧醱酵產氫試驗 60
4-2-3廚餘厭氧醱酵產氫系統出流水分析 62
4-3利用廚餘厭氧醱酵出流水結合光合連續流產氫試驗 64
4-4紫色不含硫光合細菌與藍綠細菌共培養於產氫系統中可行性試驗 67
4-4-1紫色不含硫光合細菌與藍綠細菌共培養最佳產氫混合比試驗 67
4-4-1-1單獨紫色不含硫光合細菌於共培養系統產氫試驗 67
4-4-1-2 單獨藍綠細菌於共培養系統產氫試驗 69
4-4-1-3紫色不含硫光合細菌與藍綠細菌混合比例為1：1於共培養系統產氫試驗 70
4-4-1-4紫色不含硫光合細菌與藍綠細菌混合比例為2：1於共培養系統產氫試驗 73
4-4-1-5紫色不含硫光合細菌與藍綠細菌混合比例為1：2於共培養系統產氫試驗 76
4-4-1-6紫色不含硫光合細菌與藍綠細菌混合比例為3：1於共培養系統產氫試驗 78
4-4-1-7紫色不含硫光合細菌與藍綠細菌共培養於不同混合比例下之綜合結果探討 81
4-4-2紫色不含硫光合細菌與藍綠細菌混合比例為1：1於共培養系統只利用果糖做為碳源產氫試驗 86
4-4-3藍綠細菌單獨培養於共培養系統提供乙酸做為碳源利用情形 89
4-4-4紫色不含硫光合細菌WP3-5與藍綠細菌CH3混合比例為1:1時，利用糖蜜廢水做為基質之試驗 92
第五章 結論與建議 96
5-1 結論 96
5-2 建議 98
參考文獻 99

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