臺灣博碩士論文加值系統

偶氮染料為工業上使用最普遍的一種染料，全世界市場的佔有率為60-70％，至少有3000多種類的偶氮染料應用在不同的工業上用途上。雖然，偶氮染料通常不具有劇烈的毒性，但由於環境中具有高色度，故將偶氮染料視為環境污染物。目前，每年約有高達10∼15％的合成染料隨放流水流入環境中，而且藉由傳統的生物處理程序是無法有效去除廢水中的染料。因此，過去二十年來，許多物理-化學方面的脫色技術陸續發展出來，但卻很少真正地應用在工業上，最主要的原因在於成本過於昂貴。因此，能有效去除色度且符合經濟效益的新型生物處理技術愈來愈受重視。本論文之主要研究目的在評估利用流體化床反應器合併固定化菌體顆粒之組合系統進行偶氮染料分解之的可行性，分成五個主要研究主題(第2章-第7章)。
第一個部份主要研究的主題在於評估修飾的聚乙烯醇(PVA)固定化技術應用於紡織染料脫色之可行性(第二章)。實驗結果證實修飾的PVA固定化技術確實可成功應用在連續攪拌反應器中進行染料之脫色，而且PVA菌體顆粒之高強度及具有彈性的性質也顯示其亦可應用於具有高剪應力之反應器中。另外，從電子顯微鏡的菌相觀察中，發現至少有三種具有脫色能力之微生物分布在顆粒的內部及外部，例如，球菌、桿菌及絲狀菌。
第二個部份主要研究的主題是純化分離經過染料馴養的活性污泥(石化廠)及湖泊之底泥(清華大學)中之偶氮染料分解微生物，並同時探討所分離出的偶氮染料分解菌株之染料脫色機制及其影響菌株脫色的環境因子，研究內容共有二章(第三及第四章)。經過篩選分離後，本研究得到數株具有分解偶氮染料之細菌，其中Aeromonas hydrophila 是所有篩選菌株中對所測試之24種染料(azo, anthraquinone, and indigo dyes)具有最佳的脫色能力。研究結果此菌株雖在好氣或攪拌的環境中具有較佳之生長，但是卻必須在缺氧或厭氣環境中才具有脫色的能力，當生長環境中溶氧> 0.45 mg/L 時便有明顯的抑制菌株脫色作用。實驗結果也發現，當生長環境中含有葡萄糖的存在時，Aeromonas hydrophila之脫色能力會被抑制，此乃因為菌株在生長過程中，先將葡萄糖代謝分解產生有機酸，使得培養基中的pH值下降而導致了抑制菌株之生長及脫色能力。另一方面，實驗結果也證實本研究之脫色菌株之脫色程序主要是酵素還原作用以及小部份之非活性菌體細胞之吸附作用。
第三個部份主要研究的主題是偶氮染料在固定化脫色菌顆粒內之質傳問題及分解動力現象的探討(第五章)。實驗結果顯示在可以忽略外部質傳的條件下，固定化菌體顆粒內之PVA高分子濃度及菌體濃度是影響偶氮染料在顆粒內質傳之最重要的因子。此外，本研究亦發展一種方法，從量測的反應速率來求得固定化菌體顆粒固有的(intrinsic)動力學參數，此一方法及觀念可以被應用在分析其他的固定化菌體系統。本研究也估算在不同條件之微觀環境及巨觀環境下之固定化菌體顆粒的有效因子，結果顯示固定化顆粒之內部質傳對偶氮染料之分解是一個很重要的影響因素，唯有當Thiele modulus (Φ) < 0.3 (亦即顆粒粒徑 < 0.475 mm)時，有效因子才會趨近於1，而此條件下固定化菌體顆粒分解偶氮染料之反應是屬於酵素反應限制速率步驟，而顆粒之內部質傳阻力是可以忽略的。
第四個部份主要研究的主題是關於固定化菌體顆粒在液固流體化床反應器內的水力特性(第六章)。將本研究之實驗數據與文獻報告中有關利用堅硬材料之擔體在流體化床之水力特性結果作一比較，證實文獻上利用堅硬顆粒材料擔體之水力特性，例如，拖曳係數-雷諾數、上流速度-孔隙率以及膨脹床指數-雷諾數之相關方程式等均無法適用於本研究所使用之固定化菌體顆粒，因此，本研究建立上述水力特性的新相關式，可描述PVA固定化顆粒在流體化床反應器內之流體行為。此外，在一多顆粒系統實驗中證實修正(相關)因子（f(ε)）為下降顆粒雷諾數以及流體化顆粒之阿基米得數的函數，與床孔隙度有很大相關性。本研究推導出一個簡單的關係式，在阿基米得數範圍4,986至40,745之間時，可以很容易預測床孔隙度(0.5-0.9)，而此實驗結果或許可以應用在其他類似的包埋菌體之擔體材料在流體化床反應器之水力特性。
本論文最後一個部份主要的研究主題是有關於在流體化床反應器內利用固定化菌體顆粒進行偶氮染料脫色之反應器操作特性及反應器動力模式推導，並同時將固定化菌體顆粒在流體化床反應器中之水力特性（第六章）與顆粒內部之質傳阻力（第五章）考慮在反應器動力模式中（第七章）。實驗結果顯示在固定化菌體顆粒在流體化床反應器內分解偶氮染料達穩定之反應時間會隨床膨脹、反應器內填充之顆粒密度以及水力停留時間增加而減少;而菌體在反應器內之平均停留時間，即使當水力停留時間由3小時增加至24小時，也僅有稍微被影響(1014.1-1014.9 天)，顯示以PVA當固定化擔體，即使在水力停留時間變化大時，仍可以將菌體在反應器內平均停留時間之減少衝擊降至最低。此外，在流體化床反應器動力解析方面，證實當膜數(film modulus) < 1時，顆粒之內部質傳效應對偶氮染料分解之影響會遠比顆粒外部質傳阻力(薄膜阻力)來的重要。另一方面，將反應器操作在攪拌良好的條件下進行脫色試驗，當系統達穩定時，藉由比較放流水中偶氮染料濃度與反應器動力模式所估算之染料濃度，證實此反應器動力模式之正確性。而此結果亦顯示本研究推導之反應器動力模式可以應用在設計及模擬利用固定化顆粒為擔體之液固流體化床反應器。

Azo dyes are the largest class of dyes with a world market share of 60-70%. At least 3,000 different varieties of azo dyes are extensively used in industries for various purposes. Although the azo dyes do not generally display acute toxicity, they are regarded as water pollutants because of their high color intensity in the environment. Approximately 10-15% of the amount of synthetic dyes produced annually is discharged with factory aqueous effluents, and most dyes are resistant to degradation by traditional biological processes. In the past two decades, several physical or chemical decolorization techniques have been developed, but few of them are accepted by the industries. Their lack of implementation is due to a high cost. As a viable alternative, a novel biological process has received an increasing interest owing to their cost effectiveness and environmental benignity. The main aim of the research was to determine the feasibility of the combined methods using fluidized bed reactor with immobilized-cell beads in decolorization of azo dye. The description of this study is divided into five sections.
The first section is focused on the evaluation of modified polyvinyl alcohol (PVA) immobilized technique in decolorization of textile dyes (Chapter 2). The results prove that the modified PVA immobilization technique has been successfully applied to the decolorization process in a continuous stirred tank reactor. The elasticity and high mechanical strength of PVA beads was also proven to be adequate for the high shear stresses encountered in the reactor. Microscopic observation revealed that the microbial consortium contained in the gel beads was at least made up of three kinds of bacterial species, i.e., circle-shaped bacteria, rod-shaped bacteria, and filamentous bacteria.
The second section of this research, covered in two chapters (Chapter 3 and Chapter 4), deals with the isolation of the azo-degrading bacteria from the activated sludge of petrochemical plants and the mud of University Lake. The mechanism of dye degradation using the isolated strains and the factors affecting biodegradation of azo dyes were also investigated. Several bacterial strains with the capability of degrading textile dyes were isolated. Aeromonas hydrophila was selected and identified because it exhibited the greatest decolorization ability both in terms of extent and rapidity of decolorization for 24 kinds of azo, anthraquinone, and indigo dyes. Although A. hydrophila displayed good growth in aerobic or agitation culture, color removal was the best in anoxic or anaerobic culture. Dissolved oxygen (DO) concentration exceeding 0.45 mg l-1 inhibited significantly bacterial decolorization. Glucose inhibits bacterial decolorization activity because the consumed glucose was converted to organic acids that might decrease the pH of the culture medium, thus inhibiting the cell growth and decolorization activity. In addition, preliminary results indicated that decolorization proceeded primarily by enzymatic reduction associated with a minor portion of biosorption onto the inactivated microbial cells.
The third section deals with the mass transfer phenomena and the degraded dynamics of azo dyes in the PVA immobilized-isolate beads (Chapter 5). The concentration of polymer and the density of the cell in the gel beads were the most significant influencing factors on dye diffusion at a negligibly external mass-transfer resistance condition. In addition, we have developed a procedure, which permit extraction of the intrinsic kinetic parameters from observed reaction rates. This method and concept may be used to analyze the performance of other immobilized-cell systems. On the other hand, the effectiveness factors of immobilized-cell beads have been calculated for a range of microenvironmental conditions and macroenvironmental conditions. The results showed that intraparticle diffusional resistance has a significant effect on the azo dye biodegradation rate. The calculated effectiveness factor (ηcal) approached unity at Thiele modulus (Φ) < 0.3 (dp < 0.475 mm). The only limitation was believed to be the enzymatic reaction. The internal diffusional limitations were neglected.
The fourth section deals with the hydrodynamic behaviors of PVA immobilized-cell beads in the liquid-solid fluidized bed bioreactor (Chapter 6). Our new experimental results were presented and compared with published results for the drag coefficient-Reynolds number, velocity-voidage, and expansion index-Reynolds number relationships observed during fluidization of rigid particles in a fluidized bed. Predictions made from previous correlations were examined with our new experimental findings, revealing the inadequacy of most of these correlations. Thus, new correlations describing the above-mentioned relationships are suggested. For multiparticle systems, the correction factor, f(ε), was a function of the falling gel bead properties (Reynolds number) as well as the fluidized gel bead properties (Archimedes number), and depended strongly on the bed voidage (ε). A new simple relation was developed to predict easily theεvalue from 0.5 to 0.9 at 4986< Ar < 40745. The experimental results may also be used to analyze the hydrodynamic performance of other various entrapped materials in liquid-solid fluidized bed bioreactor (FBR).
The final section deals with the operational performances and modeling study of the liquid-solid FBR using immobilized-cell beads on decolorization of azo dye. The hydrodynamic characteristics of particles (Chapter 6) and the internal diffusional limitations (Chapter 5) were taken into account (Chapter 7). It was found that azo dye degradation time initially reaching a steady state decreased with an increase in bed expansion, cell bead number density and hydraulic retention time (HRT). The mean cell residence time (θC) on liquid-solid FBR using PVA immobilized-cell beads increased insignificantly from 1014.1 to 1014.9 days as the HRT increased from 3 to 24 h, and thus the impact of conventional reducedθC can be minimized by using the same polymer as support carries. Additionally, the internal mass-transfer resistance rather than the film diffusion resistance played an important role in azo dye utilization in liquid-solid FBR when film modulus (mf) was smaller than 1. On the other hand, the model was validated through the comparisons with the experimental data that the azo dye biodegradation was at a steady state under well-mixed operating conditions. The results also showed that the model was close enough to be used in the design and simulation of liquid-solid FBR operated with immobilized-cell bead system.

CONTENTS
中文摘要………………………………………………………………..I
英文摘要………………………………………………………………IV
Figure contents……………………………………………….. ..IX
Table contents……………………………………………………...XII
1 Microbial Decolorization of Textile Dyes: Introduction………………………………………………………. 1
2 Decolorization of Azo Dye Using PVA-Immobilized Microorganisms………………………………………………… 22
3 Microbial Decolorization of Azo Dyes by Proteus mirabilis……………………………………………. 54
4 Decolorization of Textile Dyes by Newly Isolated bacterial strains………………………………………. 72
5 A Method in Evaluation of Intrinsic Kinetic Parameters on Azo Dye Biodegradation Using PVA-Immobilized Cell Beads……102
6 The Hydrodynamic Characteristics of Immobilized-Cell Beads in A Liquid-Solid Fluidized Bed Bioreactor …………….140
7 An Experimental and Modeling Study of Decolorization of Azo Dye in A Liquid-Solid Fluidized Bed Bioreactor Using Immobilized-Cell Beads………………………………..171

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