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研究生： 陳姿蒨
Zih-Chien Chen
論文名稱： 以游離酵素促進土壤中氯酚化合物偶合反應之研究
Oxidative coupling of chlorophenols catalyzed by isolated enzymes
指導教授： 李俊福
Jiunn-Fwu Lee
口試委員:
學位類別： 碩士
Master
系所名稱： 工學院 - 環境工程研究所
Graduate Institute of Environmental Engineering
論文出版年： 2016
畢業學年度： 104
語文別： 中文
論文頁數： 183
中文關鍵詞： Peroxidase 、Laccase 、氯酚化合物 、催化偶合
外文關鍵詞： Peroxidase, Laccase, chlorophenols, oxidative-coupling
相關次數： 點閱：18  下載：0 
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    • 氯酚化合物具頑抗、難分解之特性，常見處理方式有物理、化學、生物等，利用酵素去除氯酚被認為對環境較為友善。本研究藉由添加酵素至受氯酚污染土壤中，將小分子具毒性之氯酚化合物偶合成分子量大且特性穩定之產物。選取市售Peroxidase及Laccase分別與氯酚於水溶液及土壤中進行催化偶合反應，探討酵素活性、外在環境因子與反應時間對氯酚化合物去除之影響，並鑑定偶合產物及其相對產量。
    研究結果發現以相同酵素活性催化偶合反應後，當土壤有機質含量越高時，土壤中氯酚剩餘濃度由高至低為：紗帽山>彰化表土>台中土>林口土。土壤含水率越高，對酵素催化偶合氯酚化合物有明顯助益。酵素催化土壤中氯酚化合物，所產生之偶合產物不盡相同，利用Laccase分別與含2,4-二氯酚及2,4,6-三氯酚之台中、紗帽山土反應所產生之偶合產物分子量皆為339.1(m/z)，但因土壤中複雜之有機、無機質交互作用而使偶合產物相對產量與添加酵素活性並無相關趨勢，但其隨催化反應時間增長，氯酚濃度下降並伴隨偶合產物產生。
    Peroxidase及Laccase催化氯酚化合物，可去除氯酚並同時產生偶合產物。添加不同酵素催化氯酚化合物時會產生多種不同之偶合產物，於酵素催化2,3,4,5-四氯酚及五氯酚水溶液會產生七種偶合產物。Peroxidase去除氯酚化合物之效率較Laccase高，酵素催化偶合同種氯酚化合物，在土壤所產生之偶合產物分子量不同於氯酚水溶液。

    Chlorophenols (CPs) pollution of the soil has become a major concern. The pollution of soil has been treated using physical and chemical processes that have proven to be much more expensive. By changing the traditional methodology, biodegradation has been used as a new methodology in which enzymes are used (peroxidase and laccase) to promote oxidative-coupling reaction between recalcitrant organic pollutants (e.g. 2,4-DCP, 2,4,6-TCP, 2,3,4,5-TeCP, PCP) and soil organic matter (SOM), which produce stable incorporated polymers.
    In this study, the soils from Shamao Mountain, Changhua topsoil, Linkou and Taichung were collected and studied. The soil organic matter (SOMs) was determined. The SOMs and reaction time will affect enzyme activity in the environment. 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and catalyzing in aqueous medium are the optimizations to find the best activity reaction conditions with enzyme. Dissociated enzymes on the oxidative-coupling reaction in the difference phase (water and soil) will be discussed. The presence of microorganisms and organic matter in the soil as a coupling product , can promote the coupling reaction. The chlorine monomer (CPs in the soil) can be transfered into dimers, trimers of macromolecular products, which are stable incorporated polymer.When the content of organic matter is higher, the residual concentration of chlorophenols are also higher. Additionally, the moisture(%) of soil favors for oxidative-coupling reaction, and the effect of removal of chlorophenol is as follows; 100%>60%>30%. Relative quantitiy of coupling products is different in sterilization and unsterilization. When samples 2,4-DCP and 2,4,6-TCP of Taichung.; Shamao Mountain are catalyzed by laccase, their products which molecular weight were the same (339.1m/z). The oxidative-coupling reaction in soil is complicated so the trends can’t be correlated. Taking advantage of enough activity, enzyme can reduce chlorophenols and produce lots of coupling products in the same time. The effect of catalysts to chlorophenols is observed as that peroxidase showed better activity than laccase. Moerover, if numbers of chlorine in chlorophenols is higher, it will be more difficulty to remove.

    目錄　 I 圖目錄　V 表目錄 XIII 第一章 前言　1 1-1 研究緣起　1 1-2 研究目的及內容　2 第二章 文獻回顧　4 2-1 土壤基本性質及作用機制　4 2-1-1 土壤有機質　4 2-1-2 腐植質之形成機制　 5 2-1-3 土壤中影響有機污染物傳輸之因子　7 2-2 氯酚類化合物介紹　12 2-2-1 氯酚化合物之來源、性質及特性　12 2-2-2 國內氯酚化合物相關管制法規　14 2-2-3 氯酚化合物之毒性及危害　15 2-2-4 土壤中氯酚化合物之處理技術　15 2-2-5 水中氯酚化合物之處理技術　17 2-3 酵素(Enzyme)　21 2-3-1 特性與種類　22 2-3-2 Laccase　24 2-3-3 Peroxidase　26 2-3-4 影響土壤/地下水微生物胞外酵素活性之環境因子　27 2-3-5 酵素之應用　30 2-3-6 酵素在環工上的應用　31 2-4 偶合反應(oxidative-coupling)　32 2-4-1 傅里德－克拉夫茨反應(Friedel－Crafts reaction)　33 2-4-2 酵素於偶合作用扮演之角色　33 2-4-3 酵素於酚類偶合作用過程中可能之機制　35 第三章 實驗方法　39 3-1 實驗內容　39 3-2 實驗設備與儀器　41 3-3 實驗材料　48 3-3-1 土壤　48 3-3-2 溶劑　48 3-3-3 氯酚標準品　50 3-3-4 酵素　52 3-3-5 其他　52 3-4 實驗方法　54 3-4-1 土壤選擇　54 3-4-2 土壤性質分析與調整　56 3-4-3 酵素活性分析　60 3-4-4 不同時間酵素對氯酚水溶液催化反應之影響　61 3-4-5 不同酵素活性對氯酚水溶液催化反應之影響　61 3-4-6 不同時間酵素對氯酚土壤催化反應之影響　62 3-4-7 不同酵素活性對氯酚土壤催化反應之影響　62 第四章 結果與討論　63 4-1 氯酚水溶液之催化實驗　63 4-1-1 溶劑影響酵素活性之測試　63 4-1-2 Laccase氧化還原電位之測試　64 4-1-3 不同時間酵素對氯酚之偶合反應　65 4-1-4 不同酵素活性對氯酚去除之影響　70 4-1-5 LC-MS鑑定偶合產物　74 4-1-6 不同時間酵素對氯酚水溶液之偶合反應　81 4-1-7 不同酵素活性對氯酚水溶液之偶合反應　84 4-2 酵素催化偶合土壤中氯酚之前置實驗　88 4-2-1 索式萃取土壤中氯酚回收率之測定　88 4-2-2 土壤中氯酚於自然環境中之揮發試驗　90 4-3 不同條件下酵素催化土壤中氯酚之反應　95 4-3-1 不同時間酵素對氯酚土壤催化之影響　95 4-3-2 土壤含水率對酵素催化氯酚之影響　107 4-3-3 不同酵素活性對氯酚土壤催化反應之影響　111 4-3-4 土壤有機質含量對酵素催化之影響　118 4-4 酵素催化氯酚土壤偶合反應之產物　121 4-4-1 不同時間酵素對氯酚土壤偶合產物之影響　124 4-4-2 不同酵素活性對氯酚土壤偶合產物之影響　130 4-4-3 不同土壤含水率下酵素對氯酚土壤偶合產物之影響　136 4-4-4 不同土壤有機質含量對酵素偶合氯酚土壤之影響　137 4-4-5 LC-MS鑑定偶合產物　140 第五章 結論與建議　152 5-1 結論　152 5-2 建議　153 參考文獻　155 圖目錄 圖2-1 腐植質之有機質分解及形成　 6 圖2-2 Laccase之可能反應機制示意圖　 25 圖2-3 Laccase與基質ABTS催化反應圖　 25 圖2-4 Peroxidase之反應示意圖　26 圖2-5 受質濃度與反應速率之關係圖　28 圖2-6 傅－克反應：(1)傅－克烷基化：(2)傅－克醯基化反應　33 圖2-7 Laccase催化酚類氫氧基示意圖　34 圖2-8 Peroxidase催化酚類氫氧基示意圖　34 圖2-9 苯酚類C-O鍵及C-C鍵偶合可能之反應機制圖　 36 圖2-10 苯酚類C-O鍵偶合可能之反應機制圖　37 圖2-11 酵素催化氧化2,4-dichlorophenol生成自由基偶合可能之反應機制圖　38 圖3-1 研究流程圖 40 圖3-2 索式萃取裝置圖 45 圖3-3 減壓濃縮裝置圖 46 圖3-4 氮氣吹除裝置圖 47 圖3-5 丙酮結構圖 49 圖3-6 甲醇結構圖 49 圖3-7 2,4-二氯酚結構圖 50 圖3-8 2,4,6-三氯酚結構圖 50 圖3-9 2,3,4,5-四氯酚結 51 圖3-10 五氯酚結構圖 51 圖3-11 ABTS結構圖 52 圖3-12 氯酚污染土壤製備流程圖 59 圖3-13 酵素活性測試流程圖 60 圖3-14 不同活性酵素催化氯酚水溶液實驗流程 61 圖3-15 酵素催化氯酚污染土壤實驗流程 62 圖4-1 酵素於不同溶劑中之相對活性 63 圖4-2 不同ABTS添加量對五氯酚偶合反應之影響 64 圖4-3 Laccase偶合2,3,4,5-四氯酚產物液相層析儀圖譜 65 圖4-4 Peroxidase偶合2,3,4,5-四氯酚產物液相層析儀圖譜 66 圖4-5 Laccase偶合五氯酚產物液相層析儀圖譜 66 圖4-6 Peroxidase偶合五氯酚產物液相層析儀圖譜 66 圖4-7 Peroxidase隨不同時間催化2,3,4,5-四氯酚偶合曲線 68 圖4-8 Laccase隨不同時間催化2,3,4,5-四氯酚偶合曲線 69 圖4-9 Peroxidase隨不同時間催化五氯酚偶合曲線 69 圖4-10 Laccase隨不同時間催化五氯酚偶合曲線 70 圖4-11 不同活性Peroxidase催化2,3,4,5-四氯酚水溶液偶合曲線(反應5分鐘) 71 圖4-12 不同活性Peroxidase催化2,3,4,5-四氯酚水溶液偶合曲線(反應1小時) 71 圖4-13 不同活性Laccase催化2,3,4,5-四氯酚水溶液偶合曲線(反應1小時) 72 圖4-14 不同活性Laccase催化2,3,4,5-四氯酚水溶液偶合曲線(反應24小時) 72 圖4-15 不同活性Peroxidase催化五氯酚水溶液偶合曲線(反應5分鐘) 73 圖4-16 不同活性Laccase催化五氯酚水溶液偶合曲線(反應1小時) 73 圖4-17 2,3,4,5-四氯酚水溶液質譜圖 74 圖4-18 2,3,4,5-四氯酚水溶液MS圖 75 圖4-19 Peroxidase催化2,3,4,5-四氯酚水溶液偶合產物質譜圖 75 圖4-20 1 U/ml Peroxidase催化2,3,4,5-四氯酚水溶液 D偶合產物MS圖 76 圖4-21 1 U/ml Peroxidase催化2,3,4,5-四氯酚水溶液E偶合產物MS圖 76 圖4-22 6 U/ml Peroxidase催化2,3,4,5-四氯酚水溶液D偶合產物MS圖 77 圖4-23 6 U/ml Peroxidase催化2,3,4,5-四氯酚水溶液E偶合產物MS圖 77 圖4-24 6 U/ml Peroxidase催化2,3,4,5-四氯酚水溶液F偶合產物MS圖 78 圖4-25 Laccase催化2,3,4,5-四氯酚水溶液偶合產物質譜圖 79 圖4-26 6 U/ml Laccase催化2,3,4,5-四氯酚水溶液A偶合產物MS圖 79 圖4-27 6 U/ml Laccase催化2,3,4,5-四氯酚水溶液B偶合產物MS圖 79 圖4-28 6 U/ml Laccase催化2,3,4,5-四氯酚水溶液C偶合產物MS圖 80 圖4-29 1 U/ml Peroxidase催化五氯酚水溶液G偶合產物MS圖 80 圖4-30 1 U/ml Laccase催化五氯酚水溶液H偶合產物MS圖 81 圖4-31 Peroxidase催化2,3,4,5-四氯酚水溶液偶合產物相對產量趨勢圖 82 圖4-32 Laccase催化2,3,4,5-四氯酚水溶液偶合產物相對產量趨勢圖 82 圖4-33 Peroxidase催化五氯酚水溶液偶合產物相對產量趨勢圖 83 圖4-34 Laccase催化五氯酚水溶液偶合產物相對產量趨勢圖 83 圖4-35 Peroxidase催化2,3,4,5-四氯酚水溶液偶合產物相對產量趨勢圖(反應5分鐘) 85 圖4-36 Peroxidase催化2,3,4,5-四氯酚水溶液偶合產物相對產量趨勢圖(反應60分鐘) 85 圖4-37 Laccase催化2,3,4,5-四氯酚水溶液偶合產物相對產量趨勢圖(反應1小時) 86 圖4-38 Laccase催化2,3,4,5-四氯酚水溶液偶合產物相對產量趨勢圖(反應24小時) 86 圖4-39 Peroxidase催化五氯酚水溶液偶合產物相對產量趨勢圖(反應5分鐘) 87 圖4-40 Laccase催化五氯酚水溶液偶合產物相對產量趨勢圖(反應60分鐘) 88 圖4-41 未滅菌土壤置於加蓋血清瓶中於不同時間之2,4-二氯酚剩餘濃度 91 圖4-42 未滅菌土壤置於燒杯中於不同時間之2,4-二氯酚剩餘濃度 91 圖4-43 滅菌土壤置於加蓋血清瓶中於不同時間之2,4-二氯酚剩餘濃度 92 圖4-44 滅菌土壤置於燒杯中於不同時間之2,4-二氯酚剩餘濃度 92 圖4-45 未滅菌土壤置於加蓋血清瓶中於不同時間之2,4,6-三氯酚剩餘濃度 93 圖4-46 未滅菌土壤置於燒杯中於不同時間之2,4,6-三氯酚剩餘濃度 93 圖4-47 滅菌土壤置於加蓋血清瓶中於不同時間之2,4,6-三氯酚剩餘濃度 94 圖4-48 滅菌土壤置於燒杯中於不同時間之2,4,6-三氯酚剩餘濃度 94 圖4-49 Peroxidase催化紗帽山2,4-二氯酚污染土壤偶合曲線 95 圖4-50 Peroxidase催化紗帽山2,4,6-三氯酚污染土壤偶合曲線 96 圖4-51 Peroxidase催化紗帽山2,3,4,5-四氯酚污染土壤偶合曲線 97 圖4-52 Peroxidase催化紗帽山五氯酚污染土壤偶合曲線 97 圖4-53 Laccase催化紗帽山2,4-二氯酚污染土壤偶合曲線 99 圖4-54 Laccase催化紗帽山2,4,6-三氯酚污染土壤偶合曲線 99 圖4-55 Laccase催化紗帽山土中2,3,4,5-四氯酚偶合曲線 100 圖4-56 Laccase催化紗帽山土中五氯酚污染土壤偶合曲線 100 圖4-57 Peroxidase催化台中土中2,4-二氯酚偶合曲線 102 圖4-58 Peroxidase催化台中土中2,4,6-三氯酚偶合曲線 102 圖4-59 Peroxidase催化台中土中2,3,4,5-四氯酚偶合曲線 103 圖4-60 Peroxidase催化台中土中五氯酚偶合曲線 103 圖4-61 Laccase催化台中土中2,4-二氯酚偶合曲線 104 圖4-62 Laccase催化台中土中2,4,6-三氯酚偶合曲線 105 圖4-63 Laccase催化台中土中2,3,4,5-四氯酚偶合曲線 105 圖4-64 Laccase催化台中土中五氯酚偶合曲線 106 圖4-65 Peroxidase催化滅菌台中土中2,4 -二氯酚偶合曲線(50ppm) 108 圖4-66 Peroxidase催化未滅菌台中土中2,4 -二氯酚偶合曲線(50ppm) 108 圖4-67 Peroxidase催化滅菌台中土中2,4,6-三氯酚偶合曲線(50ppm) 109 圖4-68 Peroxidase催化未滅菌台中土中2,4,6-三氯酚偶合曲線(50ppm) 110 圖4-69 Peroxidase催化滅菌紗帽山土中2,4 –二氯酚剩餘濃度柱狀圖 112 圖4-70 Peroxidase催化滅菌紗帽山土中2,4,6 -三氯酚剩餘濃度柱狀圖 112 圖4-71 Laccase催化滅菌紗帽山土中2,4 –二氯酚剩餘濃度柱狀圖 113 圖4-72 Laccase催化滅菌紗帽山土中2,4,6 -三氯酚剩餘濃度柱狀圖 113 圖4-73 Peroxidase催化紗帽山土中2,4 –二氯酚剩餘濃度柱狀圖 114 圖4-74 Laccase催化紗帽山土中2,4 –二氯酚剩餘濃度柱狀圖 114 圖4-75 Peroxidase催化紗帽山土中2,4,6 -三氯酚剩餘濃度柱狀圖 115 圖4-76 Laccase催化紗帽山土中2,4,6 -三氯酚剩餘濃度柱狀圖 115 圖4-77 Peroxidase催化紗帽山土中2,3,4,5-四氯酚剩餘濃度柱狀圖 116 圖4-78 Laccase催化紗帽山土中2,3,4,5-四氯酚剩餘濃度柱狀圖 116 圖4-79 Peroxidase催化紗帽山土中五氯酚剩餘濃度柱狀圖 117 圖4-80 Laccase催化紗帽山土中五氯酚剩餘濃度柱狀圖 117 圖4-81 Peroxidase催化不同有機質含量土壤中2,4 –二氯酚剩餘濃度柱狀圖 118 圖4-82 Peroxidase催化不同有機質含量土壤中2,4,6-三氯酚剩餘濃度柱狀圖 119 圖4-83 Laccase催化不同有機質含量土壤中2,4 –二氯酚剩餘濃度柱狀圖 119 圖4-84 Laccase催化不同有機質含量土壤中2,4,6-三氯酚剩餘濃度柱狀圖 120 圖4-85 Peroxidase偶合紗帽山土中2,4-二氯酚之產物圖譜 121 圖4-86 Peroxidase偶合紗帽山土中2,4,6-三氯酚之產物圖譜 121 圖4-87 Peroxidase偶合彰化土中2,4-二氯酚之產物圖譜 122 圖4-88 Peroxidase偶合彰化土中2,4,6-三氯酚之產物圖譜 122 圖4-89 Laccase偶合紗帽山土中2,4-二氯酚之產物圖譜 122 圖4-90 Laccase偶合紗帽山土中2,4,6-三氯酚之產物圖譜 122 圖4-91 Laccase偶合彰化土中2,4-二氯酚之產物圖譜 123 圖4-92 Laccase偶合彰化土中2,4,6-三氯酚之產物圖譜 123 圖4-93 Peroxidase催化滅菌紗帽山土中2,4-二氯酚及2,4,6-三氯酚偶合產物相對產量趨勢圖 124 圖4-94 Peroxidase催化未滅菌紗帽山土中2,4-二氯酚及2,4,6-三氯酚偶合產物相對產量趨勢圖 125 圖4-95 Laccase催化滅菌紗帽山土中2,4-二氯酚及2,4,6-三氯酚偶合產物相對產量趨勢圖　　　　　　　　　　　　　　　　　　　　　　　　 125 圖4-96 Laccase催化未滅菌紗帽山土中2,4-二氯酚及2,4,6-三氯酚偶合產物相對產量趨勢圖 126 圖4-97 Peroxidase催化滅菌紗帽山土中2,3,4,5-四氯酚及五氯酚偶合產物相對產量趨勢圖 126 圖4-98 Peroxidase催化滅菌台中土中2,4-二氯酚及2,4,6-三氯酚偶合產物相對產量趨勢圖 127 圖4-99 Peroxidase催化未滅菌台中土中2,4-二氯酚及2,4,6-三氯酚偶合產物相對產量趨勢圖 127 圖4-100 Laccase催化滅菌台中土中2,4-二氯酚及2,4,6-三氯酚偶合產物相對產量趨勢圖 128 圖4-101 Laccase催化未滅菌台中土中2,4-二氯酚及2,4,6-三氯酚偶合產物相對產量趨勢圖 128 圖4-102 Peroxidase催化紗帽山土中2,4-二氯酚及2,4,6-三氯酚偶合產物相對產量柱狀圖 130 圖4-103 Peroxidase催化紗帽山土中2,4-二氯酚及2,4,6-三氯酚偶合產物相對產量柱狀圖 131 圖4-104 Laccase催化紗帽山土中2,4-二氯酚及2,4,6-三氯酚偶合產物相對產量柱狀圖 131 圖4-105 Laccase催化紗帽山土中2,4-二氯酚及2,4,6-三氯酚偶合產物相對產量柱狀圖 132 圖4-106 Peroxidase催化紗帽山土中2,3,4,5-四氯酚及五氯酚偶合產物相對產量柱狀圖 132 圖4-107 Peroxidase催化紗帽山土中2,3,4,5-四氯酚及五氯酚偶合產物相對產量柱狀圖 133 圖4-108 Peroxidase催化紗帽山土中2,3,4,5-四氯酚及五氯酚偶合產物相對產量柱狀圖 133 圖4-109 Peroxidase催化紗帽山土中2,3,4,5-四氯酚及五氯酚偶合產物相對產量柱狀圖 134 圖4-110 Laccase催化紗帽山土中2,3,4,5-四氯酚及五氯酚偶合產物相對產量柱狀圖 134 圖4-111 Laccase催化紗帽山土中2,3,4,5-四氯酚及五氯酚偶合產物相對產量柱狀圖 135 圖4-112 Laccase催化紗帽山土中2,3,4,5-四氯酚及五氯酚偶合產物相對產量柱狀圖 135 圖4-113 Laccase催化紗帽山土中2,3,4,5-四氯酚及五氯酚偶合產物相對產量柱狀圖 136 圖4-114 Peroxidase催化2,4-二氯酚土壤偶合產物相對產量柱狀圖 137 圖4-115 Peroxidase催化2,4,6-三氯酚土壤偶合產物相對產量柱狀圖 138 圖4-116 Laccase催化2,4-二氯酚土壤偶合產物相對產量柱狀圖 138 圖4-117 Laccase催化2,4,6-三氯酚土壤偶合產物相對產量柱狀圖 139 圖4-118 紗帽山2,4-二氯酚土壤未添加酵素質譜圖 140 圖4-119 紗帽山2,4-二氯酚土壤未添加酵素MS圖 141 圖4-120 紗帽山2,4-二氯酚土壤添加10U/g乾土Peroxidase偶合產物質譜圖 141 圖4-121 紗帽山2,4-二氯酚土壤添加10U/g乾土Peroxidase偶合產物MS圖 142 圖4-122 紗帽山2,4-二氯酚土壤添加30U/g乾土Laccase偶合產物質譜圖 143 圖4-123 紗帽山2,4-二氯酚土壤添加30U/g乾土Laccase偶合產物MS圖 143 圖4-124 紗帽山2,4,6-三氯酚土壤未添加酵素質譜圖 144 圖4-125 紗帽山2,4,6-三氯酚土壤未添加酵素MS圖 145 圖4-126 紗帽山2,4,6-三氯酚土壤添加5U/g乾土Peroxidase偶合產物質譜圖 145 圖4-127 紗帽山2,4,6-三氯酚土壤添加5U/g乾土Peroxidase偶合產物MS圖 146 圖4-128 紗帽山2,4,6-三氯酚土壤添加5U/g乾土Laccase偶合產物質譜圖 146 圖4-129 紗帽山2,4,6-三氯酚土壤添加5U/g乾土Laccase偶合產物MS圖 147 圖4-130 紗帽山2,4-二氯酚與2,4,6-三氯酚土壤添加20U/g乾土Peroxidase偶合產物MS圖 148 圖4-131 紗帽山2,4-二氯酚與2,4,6-三氯酚土壤添加20U/g乾土Peroxidase偶合產物MS圖 148 圖4-132 紗帽山2,4-二氯酚與2,4,6-三氯酚土壤添加20U/g乾土Laccase偶合產物MS圖 149 圖4-133 台中土2,4-二氯酚與2,4,6-三氯酚土壤添加20U/g乾土Peroxidase偶合產物MS圖 149 圖4-134 台中土2,4-二氯酚與2,4,6-三氯酚土壤添加20U/g乾土Peroxidase偶合產物MS圖 150 圖4-135 台中土2,4-二氯酚與2,4,6-三氯酚土壤添加20U/g乾土Laccase偶合產物MS圖 150 圖4-136 紗帽山2,3,4,5-四氯酚及五氯酚土壤添加20U/g乾土Peroxidase偶合產物MS圖 151 圖4-137 台中土2,3,4,5-四氯酚及五氯酚土壤添加20U/g乾土Peroxidase偶合產物MS圖 151 表 目 錄 表2-1 吸附與分佈之比較 7 表2-2 氯酚類化合物之物理化學性質 13 表2-3 氯酚污染物於地下水及土壤中之法規標準值 14 表2-4 IUBMB系統六大分類 22 表2-5 常見的偶合反應 32 表2-6 來源不同之酵素及其可催化偶合之物質 35 表3-1 高效能液相層析儀分析條件 41 表3-2 UPLC-HRMS操作條件 43 表3-3 HPLC-LRMS操作條件 44 表3-4 選用土壤一覽表 48 表3-5 選用土壤之基本性質 55 表3-6 選用土壤之pH值 56 表3-7 選用土壤之初始含水率及田間容水量 58 表4-1 偶合產物代號表 67 表4-2 配置氯酚土壤之回收率 89

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