臺灣博碩士論文加值系統

由於塑膠工業上使用的有機化學製程中的副產物，多半具有生物難分解性及毒性，造成近年來許多人工合成污染物的廢水處理問題，例如：芳香族化合物所造成的污染，其結構中的苯環物性穩定，很難被微生物所分解。
本研究嘗試以鄰苯二甲酸氫鉀(Potassium Hydrogen Phthalate, KHP)作為廢水處理模型，分別以Acinetobacter sp.及活性污泥分解不同濃度的鄰苯二甲酸氫鉀，利用特殊馴化培養基探討馴化菌種對於KHP之生物氧化性及生物分解性的影響。並以相關動力學模式探討經過馴化的菌種Acinetobacter sp.對於鄰苯二甲酸氫鉀之單一碳源基質動力學，其中KHP之生物分解指標是分別利用總有機碳(TOC)與生物需氧量(BOD)作對照分析。實驗結果發現，當以KHP加入馴化培養基為主要碳源時，純菌種對分解KHP的效果較污泥雜菌為佳，且經過馴化的菌種Acinetobacter sp.存在於鄰苯二甲酸氫鉀的臨界濃度為2100 mg C/L KHP(產生抑制現象)。
利用相關動力學模式所求得的動力學參數如下：Monod氏動力學模式求得之最大比生長率(μmax)為0.011 hr -1、飽和常數(KS)為0.201 g C/L；Michaelis-Menten動力學模式求得之最大比分解率(Vmax)為0.0592 hr -1、飽和常數(K”S)為0.133g C/L。利用修飾後Michaelis-Menten動力學模式求得之動力學參數在低濃度環境下之最大比分解率(Vmax)為0.0473 hr -1、飽和常數(K”S)為0.0742 g C/L；在高濃度環境下之最大比分解率(Vmax)為0.0931 hr -1、抑制常數(Ki)為0.67g C/L。

Various organic synthetic plastics and their derivative have been reported to seriously contaminate natural wastewater and water environments, due to their poor biodegradability and cytotoxicity in living cells. For example, aromatic compounds are frequently persistent pollutants in the wastewater that their benzene ring structures can not be easily degraded by microorganisms. In this work, we chose potassium hydrogen phthalate（KHP）as a model system and studied the culture conditions influencing the biodegradation of various concentrations of KHP in aqueous solutions by the acclimated Acinetobacter sp. and activated sludge, respectively. The experimental results showed that the degradation rate by the acclimated pure culture of Acinetobacter sp. were higher than those of the acclimated activated sludge when cells grew on KHP as sole carbon source, and the inhibition took place when KHP concentration exceeding 2100 mg C/L. Throughout the experiments, we measured the degradation rates of KHP by using the total organic carbons（TOC）and biochemical oxygen demands（BOD）as the analytical method in the liquid cultures. Finally, we have studied the microbial growth kinetics and KHP biodegradation kinetics by using Monod equation and modified Michaelis-Menten equation, respectively. According to the Monod equation in a model wastewater system, apparent KS and μmax values for the degradation of KHP by Acinetobacter sp. were 0.201 g C/L and 0.011 hr -1, respectively. According to the Michaelis-Menten equation in a model wastewater system, K”S and Vmax values for the degradation of KHP by Acinetobacter sp. were 0.133 g C/L and 0.0592 hr -1 respectively. According to the modified Michaelis-Menten equation in a model wastewater system, the apparent K”S and Vmax values for the degradation of KHP by Acinetobacter sp. at low concentration less than 1200 mg C/L were 0.0742 g C/L and 0.0473 hr -1, respectively. The apparent Ki and Vmax values for the degradation of KHP by Acinetobacter sp. at high concentration greater than 1200 mg C/L were 0.67g C/L and 0.0931 hr -1, respectively.

目 錄
中文摘要 ....................................................i
英文摘要 ...................................................ii
誌謝.........................................................iv
目錄..... ....................................................v
表目錄... ...................................................vii
圖目錄... ..................................................viii
第一章 緒論..................................................1
1.1 研究動機.............................................1
1.2 研究目的.............................................3
第二章 文獻回顧..............................................5
2.1菌種馴化..............................................5
2.2生物分解之基本觀念與介紹..............................7
2.2.1 生物分解途徑...........................................8
2.2.2 一般生物分解名詞之定義................................18
2.2.3 生物分解性測試方法....................................19
2.3利用Acinetobacter屬菌種分解芳香族成分文獻回顧.............21
2.4鄰苯二甲酸氫鉀生物代謝路徑推導............................29
2.5生物分解法實驗-TOC、BDO...................................41
第三章 實驗設備、方法與步驟.................................44
3.1實驗菌種與器材............................................44
3.2實驗藥品..................................................47
3.3實驗方法與步驟............................................49
3.3.1 菌種活化..............................................50
3.3.2 菌種馴化..............................................51
3.3.3 菌種前培養............................................55
3.3.4 TOC實驗...............................................56
3.3.5 BOD實驗...............................................57
3.3.6單一基質之相關動力學模式探討............................58
3.4實驗儀器操作..............................................62
第四章 結果與討論...........................................64
4.1菌種活化生理特性..........................................64
4.2菌種馴化之必要性..........................................68
4.3馴化後菌種與活性污泥雜菌之生物分解能力比較................69
4.3.1 馴化後Acinetobacter sp.前培養實驗.....................69
4.3.2 馴化後的Acinetobacter sp.與活性污泥雜菌的生物分解
性比較................................................70
4.3.3 馴化後的Acinetobacter sp.與活性污泥雜菌的生物氧化
性比較................................................76
4.3.4 UV生物分解測定........................................79
4.4單一基質之相關動力學模式探討..............................85
第五章 結論與建議 ...........................................91
5.1結論......................................................91
5.2建議......................................................93
參考文獻.....................................................94
附錄
A 各種極限生物分解測試法之比較.........................101
B TOC1010型例行操作步驟................................102
C 乾菌重v.s OD分析檢量線...............................103
D 以Monod模式配合Lineweaver-Burk法求比生長速率動力學
參數.................................................104
E Michaelis-Menten模式配合Lineweaver-Burk法求比分解速率
動力學參數..........................................107
F 修飾後Michaelis-Menten模式配合Lineweaver-Burk法求比分
解速率動力學參數....................................110
表目錄
表2.1 水中有機物含量分析法的優缺點..........................20
表3.1 實驗設備..............................................45
表3.2 實驗器材..............................................46
表3.3 實驗藥品..............................................47
表3.4 菌種中心之活化培養養..................................50
表3.5 活性污泥之活化培養養..................................50
表3.6 第一階段馴化培養基....................................52
表3.7 第二階段馴化培養基....................................52
表3.8 第三階段馴化培養基....................................52
表3.9 BOD基鹽...............................................53
表4.1 Acinetobacter sp.利用鄰苯二甲酸氫鉀之動力學參數.......90
圖目錄
圖1.1 鄰苯二甲酸酯反應圖....................................2
圖1.2 鄰苯二甲酸酯衍生物....................................2
圖2.1 在初步階段菌種將各種芳香族與芳香族碳水化合物進行礦化作用，代謝轉變protocatechuate..................................12
圖2.2 在初步階段菌種將各種芳香族與芳香族碳水化合物進行礦化作用，代謝轉變成catechol.......................................13
圖2.3 β-ketoadipate路徑階段性控制反應步驟.................14
圖2.4 推測真菌類微生物將β-carboxy cis , cis-muconate轉變成
β-ketoadipate的分解路徑......................................15
圖2.5 單一主要反應代謝分歧機制：利用微生物代謝L-tryptophan的二次礦化路徑.................................................16
圖2.6 利用meta-與ortho-環裂解路徑形成protocatechuate與catechnol的礦化作用，藉由dioxygenase裂解二酚基團的每一階段測量...........................................................17
圖2.7 芳香族化合物的代謝利用機制...........................24
圖2.8 利用各種不同的Acinetobacter屬菌種對於七種芳香族化合物分
解的相關文獻圖...............................................25
圖2.9 利用各種不同的Acinetobacter屬菌種將芳香族化合物轉變成
Catechol.....................................................26
圖2.10 利用各種不同的Acinetobacter屬菌種將芳香族化合物轉變
成Protocatechuate............................................27
圖2.11 Acinetobacter屬菌種的Mandelate 路徑..................28
圖2.12 利用微生物分解鄰苯二甲酸氫鉀之代謝機制示意圖.........32
圖2.13 苯甲酸化合物之代謝機制...............................33
圖2.14 Pseudo putida F1之甲苯代謝路徑.......................34
圖2.15 Pseudo putida之ortho-與meta-代謝路徑.................35
圖2.16 β-ketoadipate 代謝機制...............................36
圖2.17 苯甲酸代謝與TCA循環..................................37
圖2.18 經由二羥苯的芳香化合物代謝途徑.......................38
圖2.19 Pseudo wcnas Ovals S-5氧化gentisic Acid..............39
圖2.20 鄰苯二甲酸氫鉀之可能生物代謝路徑.....................40
圖2.21 二氧化碳之生物分解性測試系統圖.......................43
圖3.1 生物分解實驗總流程圖.................................49
圖3.2 生物分解系統示意圖（巨觀上)..........................53
圖3.3 生物分解系統示意圖（微觀上）.........................54
圖3.4 菌種前培養流程圖.....................................55
圖3.5 生物分解性（TOC）實驗流程圖..........................56
圖3.6 生物氧化性（BOD）實驗流程圖..........................57
圖4.1 未經馴化的A. calcoaceticus RAG-1在菌種中心培養基環境下
生長曲線變化情形.............................................64
圖4.2 未經馴化的A. calcoaceticus RAG-1在菌種中心培養基環境下
p H變化情形..................................................65
圖4.3 未經馴化的A. calcoaceticus RAG-1在單一碳源KHP環境下的生
物分解能力之探討.............................................65
圖4.4 未經馴化的A. calcoaceticus RAG-1在不同濃度KHP環境下生長
曲線變化情形.................................................66
圖4.5 未經馴化的A. calcoaceticus RAG-1在不同濃度KHP環境下pH
值變化情形...................................................66
圖4.6 馴化與未馴化之Acinetobacter屬菌種在50 ppmC KHP環境下的
生物分解能力之探討...........................................68
圖4.7 馴化後Acinetobacter屬菌種在單一碳源的50ppmC KHP環境下
進行前培養實驗之生長曲線變化情形與生物分解能力之探討.........69
圖4.8 馴化後Acinetobacter屬菌種在單一碳源的50ppmC KHP環境下
進行前培養實驗之pH值變化情形.................................70
圖4.9 馴化後Acinetobacter屬菌種在單一碳源100ppmC KHP環境下
生長曲線變化情形.............................................71
圖4.10 馴化後Acinetobacter屬菌種在單一碳源200~600ppmC KHP環境
下生長曲線變化情形...........................................71
圖4.11 馴化後Acinetobacter屬菌種在單一碳源100ppmC KHP~600
ppmC KHP環境下pH值變化情形...................................72
圖4.12 馴化後Acinetobacter屬菌種在單一碳源100ppmC KHP~600
ppmC KHP環境下生物分解性之探討...............................73
圖4.13 活性污泥雜菌在單一碳源50~500ppmC KHP環境下生長曲線變
化情形.......................................................74
圖4.14 活性污泥雜菌在單一碳源50~500ppmC KHP環境下pH值變化
情形.........................................................74
圖4.15 活性污泥雜菌在單一碳源50ppmC KHP環境下生物分解性之探討...........................................................75
圖4.16 活性污泥雜菌在單一碳源100ppmC KHP~500ppmC KHP環境下
生物分解性之探討.............................................75
圖4.17 馴化後Acinetobacter屬菌種在單一碳源50~500ppmC KHP環境
下溶氧量曲線情形.............................................76
圖4.18 馴化後Acinetobacter屬菌種在單一碳源50~500ppmC KHP環境
下BOD曲線變化情形............................................77
圖4.19 馴化後Acinetobacter屬菌種與活性污泥雜菌在單一碳源50
ppmC KHP環境下DO與BOD曲線變化情形............................78
圖4.20 馴化後Acinetobacter屬菌種與活性污泥雜菌在單一碳源500
ppmC KHP環境下DO與BOD曲線變化情形............................78
圖4.21 單一基質10 ppmC KHP之UV圖譜..........................79
圖4.22 馴化後Acinetobacter屬菌種在單一碳源的400ppmC KHP環境
下之生長曲線變化與KHP濃度變化情形............................80
圖4.23 馴化後Acinetobacter屬菌種在單一碳源400ppmC KHP環境下
之菌種生長前後之UV圖譜變化情形...............................81
圖4.24 馴化後Acinetobacter屬菌種在單一碳源的1400ppmC KHP環境
下之生長曲線變化與KHP濃度變化情形............................82
圖4.25 馴化後Acinetobacter屬菌種在單一碳源1400ppmC KHP環境下
之菌種生長前後之UV圖譜變化情形...............................82
圖4.26 馴化後Acinetobacter屬菌種在單一碳源的2400ppmC KHP環境
下之生長曲線變化與KHP濃度變化情形............................83
圖4.27 馴化後Acinetobacter屬菌種在單一碳源2400ppmC KHP環境下
之菌種生長前後之UV圖譜變化情形...............................83
圖4.28 不動桿菌在不同濃度KHP環境下的單一基質生長動力學
.............................................................85
圖4.29 不動桿菌在不同濃度KHP環境下的單一基質分解動力學
.............................................................86
圖4.30 馴化後Acinetobacter屬菌種在單一碳源1200ppmCKHP~1700
ppmC KHP環境下生長曲線變化情形..............................87
圖4.31 馴化後Acinetobacter屬菌種在單一碳源1800ppmC KHP~2300
ppmC KHP環境下生長曲線變化情形...............................87
圖4.32 馴化後Acinetobacter屬菌種在單一碳源1200ppmC KHP~1700
ppmC KHP環境下生物分解性之探討...............................88
圖4.33 馴化後Acinetobacter屬菌種在單一碳源1800ppmC KHP~2300
ppmC KHP環境下生物分解性之探討...............................89

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