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研究生:陳芳草
研究生(外文):Phuong-Thao Tran
論文名稱:利用固定化細胞降解MethylOrange染料
論文名稱(外文):Biodegradation of Methyl Orange by Suspended and Immobilized Pseudomonas putida mt2
指導教授:莊瑞鑫莊瑞鑫引用關係
指導教授(外文):Ruey-Shin Juang
學位類別:碩士
校院名稱:元智大學
系所名稱:化學工程與材料科學學系
學門:工程學門
學類:綜合工程學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:英文
論文頁數:67
中文關鍵詞:生物降解固定化
外文關鍵詞:BiodegradationImmobilization
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利用固定化細胞降解Methyl Orange 染料
Azo dyes which are considered to be the most recalcitrant and persistent among all groups of dye were biodegraded by various kinds of bacteria. In this study, the decolorization of Methyl Orange was determined under different conditions by non-immobilized and immobilized Pseudomonas putida mt2.
For non-immobilized cell system, it was further confirmed that decolorization was much more favorable under anoxic condition since no dye degradation was obtained with 200 rpm shaking, whereas 100% dye was removed in static condition after 3 incubation days. Temperature and pH dependences were evaluated based on the specific decolorization rate and the equilibrium conversion values to investigate the highest capability of Pseudomonas putida mt2 for decolorization in static condition. The optimal temperature range is quite narrow (33oC to 35oC) and the decolorization seems not to be suitable in acidic medium since the optimal pH for methyl orange decolorization occurred at pH 7.0 and significantly decreased at pH 5.0. The Michalis Menten equation was utilized to establish the dependence of the specific decolorization rate on the concentration of dye. The kinetic parameters of Vmax and Km were predicted up to 7.5 mg g-1 h-1 and 283 mg L-1, respectively. External diffusion coefficient kL was evaluated 7.5 x 10-6 cm s-1.
The Ca-alginate immobilized cells can not improve aerobic degradation when the dye color decreased insignificantly but completely disappeared under anaerobic condition. The optimal range of pH and temperature were obtained at 7 to 9 and 35oC to 37oC, respectively. The effects of initial biomass and initial dye concentration were also determined to confirm the predominance of immobilized cells on dye treatment when comparing with free suspended cells. Kinetics parameters were also determined to give the values of Vmax of 6.3 mg g-1 h-1 and Km of 257 mg L-1. The internal diffusion coefficient inside the beads was also investigated to give the value of 6.2 x 10-5 cm2 s-1.
Paint-PVA biofilm was created to immobilize cells instead of Ca-alginate beads when the integrity of beads was not fully maintained. Since the degradation took very long duration, the procedure of biofilm preparation was tried to recover cell’s activity. Better expression was obtained with twice adaptation. Paint-PVA immobilized cells showed the best performance on biodegradation at 35oC to 37oC, the identical optimal temperature rang with Ca-alginate immobilized cells, but the favorable range of pH is quite large from 5 to 9. In this system, the kinetics was also done to obtain the values of Vmax of 2.66 mg g-1 h-1 and Km of 161 mg L-1. The slow biodegradation rate was explained by small value of diffusion coefficient of 2.12 x 10-7 cm s-1.
Acknowledgement III
Table of contents IV
List of figures VII
List of tables IX
Chapter 1: Introduction 1
1.1 Azo dye and wastewater problem 1
1.2. Biodegradation and treatment of azo dye 2
1.2.1. Physical and Chemical treatment 2
1.2.2. Biodegradation 2
1.2.3. Mechanism of biodegradation of Methyl Orange 3
1.3. Growth patterns in batch culture 4
1.4. Biodegradation kinetics by non-growth organisms 7
1.5. Immobilization Methods 10
1.5.1. Sodium alginate 10
1.5.2. Paint and polyvinyl alcohol 11
1.5.3. Entrapment 12
1.6. Motivation and objective 12
Chapter 2: Materials and instruments 15
2.1. Chemical reagents 15
2.2. Instruments 15
2.3. Microorganism and nutrient medium 16
Chapter 3: Decolorization of methyl orange by 17
3.1. Introduction 17
3.2. Methods 18
3.2.1. Biomass and culture condition 18
3.2.2. Decolorization assay 18
3.3. Results and discussion 18
3.3.1. Cell growth curve 18
3.3.2. Dissolved oxygen influence on decolorization 20
3.3.3. Cell ages effects 21
3.3.4 Cell mass effects 22
3.3.5 Optimal pH 22
3.3.6. Optimal temperature 23
3.3.7. Dye concentration effects 24
3.3.8. Kinetics for degradation 27
3.4. Conclusions 30
Chapter 4: Decolorization of Methyl Orange by 32
4.1. Introduction 32
4.2. Methods 32
4.2.1 Cell immobilization 32
4.2.2. Decolorization 33
4.2.3 SEM study 34
4.3. Results and discussion 34
4.3.1. Shake and static comparison 34
4.3.2. Optimal pH and temperature 35
4.3.3. Effect of initial biomass on immobilized efficiency and decolorization 36
4.3.4. Decolorization kinetics for Ca-alginate immobilized system 38
4.3.5. Morphology of Ca-alginate immobilized cell 41
4.4. Conclusions 42
Chapter 5: Dye degradation by paint-PVA biofilm 43
5.1 Introduction 43
5.2. Methods 43
5.2.1. Biofilm preparation 43
5.2.2. Decolorization assay 43
5.3. Results and discussions 44
5.3.1. Improvement of degradation rate 44
5.3.2. Large spectrum of favorable pH 44
5.3.3. Optimal temperature 45
5.3.4. Dye concentration effects of paint-PVA immobilized cells 46
5.3.5. Cell reuse 49
5.4. Conclusion 50
Chapter 6: Mass transfer in non-immobilized and 51
6.1. Introduction 51
6.1.1. Diffusion effects on nonporous support material 51
6.1.2. Diffusion effects in a porous matrix 53
6.2. Mass transfer and liquid diffusion coefficient in non-immobilized system 55
6.3. Mass transfer and effective diffusivity of Methyl orange within the porous matrix 58
6.4 Mass transfer and diffusion coefficient through paint-PVA biofilm 59
6.5. Discussion about mass transfer effect 60
Chapter 7: Conclusions 62
References 64
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