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系統識別號 U0026-0307201310535000
論文名稱(中文) 以藍綠菌嗜熱性聚球藻Thermosynechococcus sp. CL-1 固碳與其類胡蘿蔔素產率分析之研究
論文名稱(英文) CO2-Fixation by a Cyanobacterial Strain,Thermosynechococcus sp. CL-1, and Analysis of Its Carotenoids Productivity
校院名稱 成功大學
系所名稱(中) 地球科學系碩博士班
系所名稱(英) Department of Earth Sciences
學年度 101
學期 2
出版年 102
研究生(中文) 闕凡瑜
研究生(英文) Fan-Yu Chueh
學號 L46004123
學位類別 碩士
語文別 中文
論文頁數 135頁
口試委員 指導教授-羅尚德
共同指導教授-朱信
口試委員-吳建一
中文關鍵字 二氧化碳  薄板反應器  類胡蘿蔔素  藍綠菌 
英文關鍵字 CO2 fixation  flat plate photobioreactors  carotenoids  cyanobacteria  Thermosynechococcus sp. 
學科別分類
中文摘要 近年來,全世界人口快速增加和化石燃料的大量使用,導致大氣中二氧化碳濃度上升,因而產生溫室效應、全球暖化等氣候異常問題,因此管制二氧化碳排放量並同時發展二氧化碳減量技術為世界各國永續發展所需達成的目標。
藍綠菌具有高固碳速率和環境耐受性佳之特性,各國均效力於開發微藻(或藍綠菌)固碳技術並將太陽能轉換成化學能,因此,為使藍綠菌固定二氧化碳率最大化,可採用薄板反應器在最佳生質體密度、最佳光照下大幅提升藍綠菌產率及固碳率。固碳後的的生質體可被用來生產能源或高經濟價值產品(如類胡蘿蔔素)以降低固碳所需花費之成本,增加未來實廠化的可行性。類胡蘿蔔素普遍存在於藍綠菌生質體內,對人體健康而言,具有延緩老化及其他慢性疾病的發生,人類無法自行合成類胡蘿蔔素且只能從飲食中攝取。因此,以藍綠菌在固碳後生產類胡蘿蔔素應用於人類飲食、醫療環境在經濟、與人類永續發展上,極具技術開發之必要性。
光照強度、鹽度為影響類胡蘿蔔素含量與藍綠菌生產之關鍵因素。此外,為了將來應用於戶外培養,須了解晝夜變化(有光照/無光照)對於類胡蘿蔔素生產之影響性,本研究以生長速率極高且耐高溫(40-62℃)、高鹼性(pH=9-12)環境之藍綠菌Thermosynechococcus sp. CL-1 (TCL-1)作為固定二氧化碳之光合菌種。以薄板反應器於批次培養(batch operation)下,探討TCL-1在不同 LED 光強度(200-1,200 μE /m2/ s)、鹽度(NaCl為0.03 M、0.25 M和0.5 M)以及LED亮/暗比(12小時:12小時;用於模擬自然界晝夜週期變化)條件下,藍綠菌之細胞生長速度、固碳速率以及類胡蘿蔔素含量與產量之影響。
研究結果發現,TCL-1在 LED 光強度為600 μE /m2/ s時細胞具有最大比生長速率、產率、固碳速率及類胡蘿蔔素產率(4.12/d、1.51 g/L/d、2.40 g/L/d和14.1 mg/L/d);1,000 μE /m2/ s時單位細胞內類胡蘿蔔素含量最高(1.09 %),而在1,200 μE /m2/ s時則會因過量光照對細胞生長造成抑制。利用不同光照條件(200、600及1,000 μE /m2/ s)進一步針對LED 亮/暗比(12小時:12小時)之情況進行探討,結果顯示在1,000μE /m2/ s條件下有其最小值,其比生長速率、產率、固碳速率和類胡蘿蔔素產率分別降低到1.43/d、0.11 g/L/d、0.15 g/L/d 和4.2 mg/L/d。藻體固碳後可萃取內含之類胡蘿蔔素,以降低固碳及培養成本。總結來說,本研究成果應能提供未來評估藻類系統應用於降低二氧化碳排放與生產具商業化效益產品(類胡蘿蔔素)之可行性。
英文摘要 Global warming and climate change have become worse due to the increased population and the excess utilization of fossil fuels. The development of CO2 sequestration technologies as well as restrain the CO2 emissions are extremely important purposes for sustainable development of human beings.
Because the cyanobacteria has the ability to fix carbon in high efficiency and the high environmental tolerability characteristics, various countries put their efforts on the development of CO2 fixation technologies for cyanobacteria (or microalgae). In addition to optimize the carbon fixation efficiency of cyanobacteria, the flat plate bio-reactor is chosen to raise the biomass productivity in the optimize biomass concentration and light intensity. After the carbon fixation, the biomass can be use as energy production precursor or high value products, such as carotenoids. However, the cost of CO2 fixation by cyanobacteria and energy production are too high. Valuable products, such as carotenoids, have high potential to reduce the cost. Carotenoids have high economic benefit which can be commonly extracted from cyanobacteria. Carotenoids have ability to decelerate aging and prevent from chronic disease for human beings. Human beings can’t produce carotenoids by themselves but only from the diet. Therefore, in order to fit the demands of CO2 fixation, healthy products and sustainable development, cyanobacteria Thermosynechococcus CL-1 (TCL-1) is cultivated in the flat plate photobioreactors to enhance the CO2 fixation rate and the biomass productivity in this study.
Light intensity and salinity are the key factors that influence carotenoids content and the productivity for cyanobacteria. In addition, for the sake of outdoor cultivation in the future, the diurnal cycle (light / dark) affect the production of carotenoids is also needs to research. In this study, thermophilic (40-62 ℃) and basophilic (pH = 9-12) cyanobacteria Thermosynechococcus sp. CL-1 (TCL-1) is chosen as the fixed carbon dioxide photosynthetic bacteria. In batch operation, TCL-1 in LED light intensity (200-1,200 μE/m2/s), salinity (NaCl 0.03 M、0.25 M and 0.5 M), the LED light / dark (12 hours: 12 hours; used to simulate the natural changes in the circadian cycle), the cyanobacteria cell growth, carbon fixation efficiency and carotenoids content, cell productivity and cost analysis are discussed. The results indicate that the TCL-1 has the maximum specific growth rate、productivity、carbon fixation rate and carotenoids productivity (4.12/d、1.51 g/L/d、2.40 g/L/d and 14.1 mg/L/d, respectively) in the case of LED light intensity of 600μE/m2/s. Cells growth inhibition occurs in the condition of 1,200μE/m2/s due to excessive light. Moreover, salt (NaCl) and LED lights / dark (12:12) was further explored by using different light conditions (200, 600 and 1,000μE/m2/s). The results show that the specific growth rate、productivity、rates of carbon fixation and carotenoids could be reduced to 1.43/d、0.11 g/L/d、0.15 g/L/d and 4.2 mg/L/d, respectively.
The knowledge and outcome obtained from this study would be useful for assessing the feasibility of using microalgae system in practical applications for CO2 mitigation and commercial carotenoids productions.
論文目次 摘要 ……………………………………………………………………………I
中文摘要 I
Abstract III
誌謝………………………………………………………………………………...Ⅴ
目錄 …………………………………………………………………………..A
表目錄 …………………………………………………………………………...E
圖目錄 …………………………………………………………………………..G
第一章 前言 1
1-1 研究動機 1
1-2 研究目的 4
第二章 文獻回顧 6
2-1 溫室效應 6
2-1.1 溫室氣體 7
2-2 二氧化碳處理方式介紹 8
2-2.1 物理處理法 9
2-2.2 化學處理法 10
2-2.3 生物固定法 13
2-3 藻類簡介 16
2-4 類胡蘿蔔素簡介 20
2-4.1 類胡蘿蔔素來源及特性 20
2-4.2 類胡蘿蔔素結構 21
2-4.3 類胡蘿蔔素之應用 23
2-5 光合作用 24
2-6 環境因子對藻類細胞之影響 30
2-6.1 培養系統 30
2-6.2 生長曲線及測定 31
2-6.3 生物培養反應器 33
2-6.4 碳、氮源的影響 37
2-6.5 光源 38
2-6.6 光照強度 40
2-6.7 鹽度 40
2-6.8 二氧化碳: 41
2-6.9 溶氧濃度: 41
2-6.10 溫度與pH值 42
2-7 發光二極體 43
2-8 細胞破碎方法簡介 47
2-8.1 細胞破碎 47
2-8.2 球磨機 48
2-8.3 超音波 48
2-8.4 熱裂解 49
第三章 材料與方法 50
3-1 研究架構 50
3-2 實驗藥品、器材及分析設備 51
3-2.1 培養基藥品 51
3-2.2 實驗器材 52
3-2.3 分析設備 54
3-3 實驗流程 55
3-4 藻株TCL-1簡介 56
3-4.1 藻株來源 56
3-4.2 藻株保存 58
3-4.2.1 低溫保種 58
3-4.2.2 凍乾保種 58
3-4.2.3 一般保種 58
3-4.3 藻株TCL-1之特性及型態 59
3-4.4 藻株TCL-1之吸收特性波長之選擇 60
3-5 反應器設計及實驗操作 61
3-5.1 生長曲線及檢量線測定 65
3-6 藻株培養流程 66
3-6.1 連續流活性培養 67
3-6.2 批次培養 70
3-6.3 藻株生長曲線測定流程 70
3-6.4 細胞最大比生長速率和細胞產率測量 71
3-7 細胞組成分析 71
3-7.1 細胞中元素分析方法 71
3-7.2 類胡蘿蔔素含量分析方法 72
第四章 結果與討論 75
4-1 光照強度對TCL-1之比生長速率、細胞產率、固碳速率及類胡蘿蔔素產率影響 75
4-1.1 比生長速率、細胞產率及pH值 75
4-1.2 固碳速率 81
4-1.3 類胡蘿蔔素產率 84
4-2 鹽度對TCL-1之比生長速率、細胞產率、固碳速率及類胡蘿蔔素產率的影響 87
4-2.1 比生長速率、細胞產率及pH值 87
4-2.2 固碳速率 93
4-2.3 類胡蘿蔔素產率 95
4-3 亮暗比(12:12)對TCL-1之比生長速率、細胞產率、固碳速率及類胡蘿蔔素產率的影響 98
4-3.1 比生長速率、細胞產率及pH值 99
4-3.2 固碳速率 105
4-3.3 類胡蘿蔔素產率 106
第五章 結論與建議 109
5-1 結論 109
5-2 建議 111
參考文獻 ………………………………………………………………………...112
附錄……………………………………………………………………….………134
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