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研究生：	薛欣達 Hsueh, Hsin-ta
論文名稱：	以光合微生物固定高溫高鹼度吸收液中碳源之研究 Bio-fixation of Carbon Source from Absorbed Solutions at High Temperature and Alkaline Conditions with Photosynthetic Microorganisms
指導教授：	朱信 Chu, Hsin 余世宗 Yu, Shih-tsung
學位類別：	博士 Doctor
系所名稱：	工學院 - 環境工程學系 Department of Environmental Engineering
論文出版年：	2007
畢業學年度：	95
語文別：	英文
論文頁數：	122
中文關鍵詞：	擬球藻、二氧化碳、質量傳輸、鹼液吸收、藍綠菌、生質能
外文關鍵詞：	Algae, Biofuel, IR-spectroscopy, 16S rDNA, Cyanobacterium, Alkaline solution, Mass transfer, Carbon dioxide
相關次數：	點閱：94 下載：4
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隨著全球暖化問題日益嚴重，溫室氣體之減除已是當務之急。其中又以最大宗之二氧化碳工業排放為首要。光合作用可固定二氧化碳又可將其轉化為生質能，十分符合能源短缺之現今。而其中光合微生物之光合效率又較陸生植物為高。
本研究首先針對二氧化碳直接曝氣培養光合微生物與模擬鹼液吸收二氧化碳之碳酸鈉添加液來培養，進行固碳效能之比較。採用之光合微生物為東港水產試驗所之Nannochloropsis oculta (NAO)及由台東金崙溫泉所篩選之hot spring cyanobacterium (HSC)。結果顯示模擬鹼液吸收二氧化碳之培養較好，且HSC有高於NAO之固碳效率。進一步由型態及分子生物鑑定後，發現HSC相近於Thermosynechococcus elongates BP-1。故命名為Thermosynechococcus sp. CL-1 (TCL-1)。
由碳源濃度對TCL-1生長速率可推導出Monod方程式。最大生長速率 (μmax) 為 3.85 ± 0.07 d-1及親和力常數 (KS) 為 1.95 ± 0.28 mM (DIC，dissolve inorganic carbon)。與煙道氣吸收液溫度十分相當之培養液溫度之實驗結果可得耗碳速率(θ)為1.02 (介於溫度40至55C之間)。
進一步，本研究採用填充塔進行實際鹼液吸收，吸收液再進行TCL-1之培養。結果發現TCL-1可耐鹼之條件下進行操作，整體除碳效能可提昇五倍之多。另外，在固定pH值(7至11)之操作下，TCL-1皆可維持1.5 d-1以上之生長速率。此系統以此耐鹼性光合微生物(TCL-1)進行二氧化碳之處理，應可有效提昇二氧化碳處理效率。
本研究亦針對不同固碳條件下所生成之微生物體(biomass)，進行其組成差異之探討。結果顯示，將TCL-1培養在中性時(pH=7)，脂質及蛋白質之含量較其它酸鹼值培養下為高。至pH 10.5時，碳水化合物為最高。但至pH 為11時，則傾向於重金屬之吸收，相同情形亦發生於DIC濃度小於18.9 mM時。大於此一濃度，脂質之生成減少而轉成為碳水化合物。另外，在不同操作條件下，生成之biomass亦有不同之裂解特性。此一結果可由熱重分析儀及紅外線分析儀之分析得到。

Carbon dioxide mass transfer is a key factor in cultivating photosynthetic microorganisms besides the light limitation of photosynthesis. Firstly, this study offers a comparison of CO2 assimilation with photosynthesis via combining with and without alkaline absorption. The two photosynthetic microorganisms adopted as the test samples were Nannochloropsis oculta (NAO) and hot spring cyanobacterium (HSC). HSC was isolated from an alkaline hot spring (pH 9.3, 62°C), Chin-Lun hot spring, in eastern Taiwan and grows well over pH 11.5 and 50°C. The growth of NAO and HSC was better when combined with alkaline absorption than without it. The integration of alkaline absorption and photosynthetic bio-fixation provides a higher performance for CO2 assimilation than photosynthetic bio-fixation alone. In addition, HSC on the growth and alkaline adaptation were better than NAO under suitable temperature and pH. After analysis of HSC phylogeny with 16S rDNA and its morphological characteristics, the species is found to be close to Thermosynechococcus elongates BP-1 in the bootstrap tree. This strain is named as Thermosynechococcus sp. CL-1 (TCL-1).
In order to assess the available of TCL-1 on the caboxylation, a Monod equation of TCL-1 with varied DIC (dissolved inorganic carbon) concentrations was proposed. The maximum growth rate (μmax) was 3.85 ± 0.07 d-1; affinity constant (KS) was 1.95 ± 0.28 mM. We also proposed an equation of CO2 assimilation rate that ranged in temperature from 40 to 55C of temperatures. The assimilation constant (θ) was 1.02.
We also performed the alkaline absorption of CO2 with a packed tower and then carboxylation with TCL-1 to do the preliminary test of integrated system of these two mechanisms, TCL-1. CO2 removal efficiencies in a packed tower increase about 5-fold in a suitable growth condition compared to that of without adding any sodium hydroxide. In addition, TCL-1 also exhibits high growth rates under the controlled pHs from 7 to 11. The integrated system is, therefore, more feasible to treat CO2 in the flue gases using the species with higher alkaline affinity such as TCL-1 in small volume bioreactors.
Alkaline conditions enhance the water’s absorption capacity of CO2, but the DIC carboxylation mechanisms under different pHs and DIC concentrations may change the composition of the biomass. Hence a study on the effects of pH and DIC regarding the content variations of four elements (C, N, H, O), lipids (LI), proteins (PR), and carbohydrates (CA) was carried out. The concentrations of PR and LI were the highest under the cultivation of pH 7 and CA was at 10.5. According to the analysis of three compositions, the production pathway of LI might be shifted to CA from pH 7 to 10.5, and shifted to inorganic compound from pH 10.5 to 11. Regarding the effect of DIC at pH 9, the results revealed that the uptake pathway shift (such as metals uptake) might happen while DIC is less than 18.9 mM. From 18.9 to 47.2 mM of DIC, the production pathway of LI shifted to CA and the contents of CA increased quickly from 47.2 to 94.3 mM without a further decrease of LI. Regarding the pyrolysis experiments with a thermogravimetric analyzer coupled with FT-IR (TG-IR), the transformation of xylan, cellulose, and lignin contents was observed under various pHs and DIC concentrations.

摘要		A
ABSTRACT	C
致謝		E
CONTENT		I
LIST OF TABLES	IV
LIST OF FIGURES	V
NOMENCLATURE	VIII
CHAPTER 1  INTRODUCTION	1
CHAPTER 2  LITERATURES REVIEW	6
1  Greenhouse effect	6
2  Technologies on the mitigation of carbon dioxide	8
3  Photosynthetic mechanism on the mitigation of carbon dioxide	10
4  Identification of cyanobacteria	18
5  Biomass (Bio-energy)	25
CHAPTER 3  EXPERIMENTAL EQUIPMENT AND PROCEDURES	29
1	Experimental reagents and equipments	29
1.1  Cultivation media and other reagents	29
1.2  Gases and accessories in the packed tower	30
1.3  Experimental equipments	30
2	Photosynthetic microorganism candidates	34
2.1  Photosynthetic microorganism sources	35
2.2  The calibration between cell density and optical density	35
3	The identification of HSC	36
3.1  Morphological identification	36
3.1.1  Observation with SEM	37
3.1.2  Observation with TEM	37
3.2  Identification with 16S rDNA	39
3.2.1  PCR	39
3.2.3  Sequence and Blast search in NCBI	41
3.2.4  Phylogenic analysis	41
4  Cultivation of photosynthetic microorganisms	45
4.1  Cultivation of NAO	45
4.1.1  Cultivation of NAO with different simulated absorbed solutions	45
4.1.2  Cultivation of NAO with various CO2 concentrations bubbling	46
4.1.3  Cultivation of NAO under various pHs and Temperatures	47
4.2  Cultivation of HSC	48
4.2.1  Cultivation of HSC with various CO2 concentrations bubbling	49
4.2.2  Cultivation of HSC under various pHs, temperatures, and DIC concentrations	49
4.2.3  The absorption of CO2 in a packed tower with NaOH and ammonia solutions	49
4.2.4  Cultivation of HSC with absorbed NaOH and ammonia solutions from the packed tower	51
5	Assessment of reuse of biomass	52
5.1  FT-IR analysis of macro-compositions of biomass	52
5.2  Wet chemistry analysis of macro-compositions of biomass	53
5.3  Element analysis	56
5.4  Metal analysis	57
5.5  Pyrolysis experiments with TGA and TG-IR	57
CHAPTER 4  RESULTS AND DISCUSSION	58
1  The preliminary test of the photosynthetic microorganisms	58
2  The identification of HSC with 16S rDNA gene and morphology	60
3  CO2 assimilation with photosynthesis integrated with or without alkaline absorption	66
3.1  The cultivation of NAO and HSC with bubbling CO2	66
3.2  The cultivation of NAO and HSC under various temperatures	71
3.3  The cultivation of NAO and HSC under various pHs	73
3.4  The cultivation of HSC under various DIC concentrations	79
3.5  The compositions (C, H, N, O and S) of NAO and HSC under various conditions	82
4  The integration test between packed tower and photo-bioreactor	82
4.1  The test of HSC	82
4.2  Screening test for the simulated absorbed solutions for marine algae	85
4.3  Sterilization methods and element additives for marine algae	88
5  Assessment of reuse of biomass	91
5.1  Elements and IR-spectroscopic quantitative analysis of biomass cultivated under various conditions	91
5.2  Pyrolysis analysis of biomass by TG-IR	101
CHAPTER 5  CONCLUSIONS AND SUGGESTIONS	107
1  Conclusions	107
2  Suggestions	109
REFERENCES	111
APPENDIX 1	117
                                    

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