臺灣博碩士論文加值系統

本研究旨在開發一空氣生物陰極式微生物燃料電池(air biocathode microbial fuel cells, AB-MFC)，並於AB-MFC之陰極植種白腐真菌─靈芝(Ganoderma lucidum BCRC 36123)以生產漆酶(laccase)，應用於提升廢水中偶氮染料之分解及評估其產電效率。本研究之動機為AB-MFC陽極槽廢水中之偶氮染料─Acid orange 7 (AO7)可藉由聚乙烯醇水凝膠(polyvinyl alcohol hydrogel, PVAH)擴散至陰極作為靈芝之碳源，除了可維持靈芝生長之外，並藉此持續分泌漆酶，作為陰極氧氣之催化劑，可提升MFC之電壓輸出。此外，也藉由PVA-H之保水與吸水性，能提供陰極保持濕潤狀態而有利於靈芝之生長，以及接收陽極端微生物降解AO7所產生之質子。研究結果顯示：(1)靈芝可以AO7作為碳源並釋出漆酶，其漆酶活性可於第19天達最大值為20.3 ± 0.2 U/L，脫色率則為77% (AO7為50 mg/L)；(2)當陰極端植種靈芝並可釋出具活性之漆酶系統(簡稱lac-WRF-AB-MFC)時，於連續添加濃度為180 mg/L之AO7作為唯一碳源時，最高開路電壓達821 mV、閉路電壓為394 mV (外電阻為1000 Ω)、最大功率密度為13.38 mW/m2、最大電流密度為33 mA/m2、脫色率則可達82%，均優於單純空氣陰極系統(簡稱AC-MFC)及白腐真菌失去漆酶活性系統(簡稱non-lac-WRF-AB-MFC)之試驗組別；(3)透過恆電位儀(potentiostat)之循環伏安法(cyclic voltammetry, CV)掃描AB-MFC系統之陰極端電位變化結果顯示，於陰極端植種靈芝並釋出漆酶時，陽極端可接收來自微生物降解過程中所遞出之電子，並透過外部電路方式傳遞至陰極，此時電子則可再透過AO7或愈創木酚(guaiacol)充當漆酶之電子傳遞介體(mediator)，讓陽極傳遞至陰極之電子轉移至漆酶活性位置(active site)中，再與陰極端氧氣反應，因此漆酶乃成為陰極端氧氣之催化劑，可帶動大氣中之氧氣快速被攜至陰極端。AB-MFC之整體研究成果顯示，空氣陰極表層生長具可釋出漆酶之靈芝時，漆酶可作為MFC系統陰極端對氧氣之催化劑，而靈芝亦可利用來自陽極端之AO7作為生長菌絲體之碳源，此產出之菌絲體可持續分泌漆酶，因此可提升整體MFC系統之電壓輸出及提高污染物去除能力。

The objective of this research is to develop an air biocathode microbial fuel cell (AB-MFC), with cultures of laccase-producing white rot fungi-Ganoderma lucidum (BCRC 36123) planted at the cathode. The developed AB-MFC was then applied to investigate the enhancement of the decomposition of azo dyes in wastewater as well as its power generation. The motivation of this study was that the azo dye acid orange 7 (AO7) in the wastewater contained within the anode chamber of the AB-MFC could diffuse to the cathode through a polyvinyl alcohol hydrogel (PVA-H), serving as a source of carbon for the Ganoderma lucidum and sustain its growth. The Ganoderma lucidum would continuously generate laccase which would serve as a catalyst for the oxygen reaction at the cathode, further improving voltage output of the MFC system. PVA-H also provided the additional functions of water retention and moisture absorption, helping to moisturize the cathode and facilitate growth of the Ganoderma lucidum as well as to receive protons generated through biodegradation of AO7 at the anode. Results of the study showed: (1) Ganoderma lucidum could utilize AO7 as a source of carbon to release laccase. AO7 concentration being 50 mg/L could be decolorized by 77% in 19 days, and the laccase activity reached a maximum of 20.3 ± 0.2 U/L. (2) The system of planting Ganoderma lucidum at the cathode to release active laccase by repeatedly adding 180 mg/L AO7 as the sole source of carbon was capable of achieving a maximum open-circuit voltage of 821 mV, maximum closed-circuit voltage of 394 mV (external resistance of 1000 Ω), maximum power density of 13.38 mW/m2, maximum current density of 33 mA/m2, and highest decolorization of 82%, all of which were superior to the air cathode system or the white-rot fungi system with inactive laccase. (3) Changes to voltage potential at the cathode of the AB-MFC system monitored using cyclic voltammetry scanning by a potentiostat revealed that planting laccase-releasing Ganoderma lucidum at the cathode allowed the anode to receive electrons generated through microbial decomposition. These electrons could then be transmitted to the cathode through external circuitry. At the cathode, these electrons could either use AO7 or guaiacol as the mediator and be delivered by the anode to the cathode to the active site of the laccase and finally reacted with oxygen. Therefore, laccase could serve as a catalyst for oxygen and accelerate the oxygen transport from atmosphere to cathode. Overall research outcomes of this AB-MFC showed that growing Ganoderma lucidum on the air cathode surface could generate laccase that catalyzes the cathode oxygen reaction of the MFC. Ganoderma lucidum could also utilize AO7 brought from anode as a carbon source for hyphal growth. The growing hyphae would generate more laccase, helping to further improve the overall voltage output and contaminant removal capabilities of the entire MFC system.

摘要 i
ABSTRACT iii
誌謝 v
目錄 vi
表目錄 x
圖目錄 xi
第一章 緒論 1
1.1 研究背景 1
1.2 研究動機 3
1.3 研究目的 4
1.4 研究內容 5
1.5 研究架構 6
第二章 文獻回顧 9
2.1 染料介紹 9
2.1.1 染料分類介紹 10
2.1.2 偶氮染料 12
2.2 染整廢水處理方法介紹 13
2.2.1 傳統染整廢水處理方式 13
2.2.2 生物處理法 15
2.3 白腐真菌 16
2.4 漆氧化酶 17
2.4.1 漆氧化酶之反應機制 18
2.4.2 漆氧化酶之應用範圍 20
2.5 微生物燃料電池 21
2.5.1 微生物燃料電池之沿革 21
2.5.2 微生物燃料電池之原理 25
2.5.3 微生物燃料電池之染整廢水應用 26
2.5.4 空氣陰極式微生物燃料電池 27
2.6 微生物燃料電池之陰極催化劑改良 28
2.7 微生物燃料電池結合漆氧化酶之應用 31
第三章 材料與方法 33
3.1 AirlacWRFACMFC組態與操作條件 33
3.2 研究設備與藥品 37
3.2.1 監測儀器 37
3.2.2 實驗藥品 37
3.3 菌種來源與空氣陰極靈芝碳布培養 39
3.3.1 菌種來源 39
3.3.2 靈芝固態培養基成分與配比 39
3.3.3 靈芝液態培養基成分與配比 40
3.3.4 空氣陰極靈芝碳布培養 40
3.3.5 初始靈芝碳布上之漆酶活性測定 42
3.3.6 PVA-H中漆酶活性測定 43
3.3.7 空氣陰極靈芝碳布之漆酶失活處理 44
3.4 漆酶相關檢測方法 45
3.4.1 漆酶固態鑑定 45
3.4.2 漆酶活性測定─ABTS檢測法 46
3.5 AO7之分析方法與脫色率計算 48
3.6 產電分析 50
3.6.1 電壓 50
3.6.2 電流 50
3.6.3 電功率 50
3.6.4 開路電壓 51
3.7 AirlacWRFACMFC系統之內電阻分析 51
3.6.1 極化曲線斜率法 51
3.6.2 功率密度峰值法 51
3.8 循環伏安法 52
第四章 結果與討論 55
4.1 白腐真菌背景試驗 55
4.1.1 漆酶固態鑑定 55
4.1.2 以AO7作為基質分泌漆酶進而自行降解AO7之脫色試驗 57
4.2 以循環伏安法證明AirlacWRFACMFC具較高氧化還原波峰之探討 58
4.2.1 基質為愈創木酚之循環伏安圖 58
4.2.2 基質為AO7之循環伏安圖 63
4.3 探討AirlacWRFACMFC系統對於空氣陰極改良之產電效率 67
4.3.1 初始靈芝碳布與PVA-H之漆酶活性測定 67
4.3.2 電壓監測 68
4.3.3 不同陰極系統之極化曲線與功率密度曲線 70
4.4 探討AirlacWRFACMFC系統對空氣陰極改良之AO7脫色效率 74
4.5 研究成果與MFC處理染整廢水文獻之相關文獻成果比較 76
第五章 結論與建議 79
5.1 結論 79
5.2 建議 80
參考文獻 81

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