油質性微藻類之培養與固體觸媒在生質柴油製程之應用

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研究生：	蘇家弘 Chia-Hung Su
論文名稱：	油質性微藻類之培養與固體觸媒在生質柴油製程之應用 Cultivation of oleaginous microalgae and the application of solid catalytic process to biodiesel production
指導教授：	吳文騰 Wen-Teng Wu 朱一民 I-Ming Chu
口試委員:
學位類別：	博士 Doctor
系所名稱：	工學院 - 化學工程學系 Department of Chemical Engineering
論文出版年：	2007
畢業學年度：	95
語文別：	中文
論文頁數：	157
中文關鍵詞：	微藻類、運動強度、三原色、兩階段培養、生質柴油、擬均相動力學模型
外文關鍵詞：	microalgae, motile intensity, primary colors, two-stage cultivation, biodiesel, pseudo-homogeneous kinetic model
相關次數：	點閱：2 下載：0
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在本研究中，我們主要進行油質性單細胞微藻類之培養系統設計與固體觸媒在生質柴油製程之應用。在藻類培養系統設計方面，由於藻類運動強度正比於藻類的比生長速率（R2=0.82），因此藻類運動強度可做為篩選藻類最適化培養基之指標。此外，藻類在培養過程中顏色的變化（生長期的深綠色至停滯期之黃綠色），被認為與藻體內部成分組成變化有關。根據這項理由，藻類顏色所發生的變化以三原色來進行分析，並與葉綠素與油脂含量作對應可得線性關係式。利用此關係式估計藻類葉綠素或油脂含量之預測值，也以實際實驗驗證其可靠性（判定係數高於0.94）。利用這樣的影像分析可以改善傳統分析方法的不便，進而協助我們快速的瞭解藻類的生理狀態以及決定藻類最佳培養策略。由於藻類生長與油脂累積的機制各自所需的條件相異，因此開發藻類兩階段培養程序以同時符合各自所需的要求。在兩階段培養策略中，第一階段培養提供富含藻類營養成分的培養環境以促進藻類大量的生長；接著再將藻類培養至第二階段培養程序內。由於第二階段為氮源缺乏的培養條件，因此藻類油脂預期會在此階段進行大量的累積。相較於傳統單一階段批次培養，此程序可獲高於2.82倍的油脂產率。
在另一方面，以大豆油做為研究藻類油脂生產生質柴油之基礎，來探討固體觸媒於生質柴油製程之應用。本研究中，先以酵素法將大豆油進行水解反應以獲得脂肪酸。在強酸性陽離子交換樹脂的催化下，水解獲得之脂肪酸與甲醇進行酯化反應生成脂肪酸甲酯。觸媒使用量、反應溫度與醇酸莫耳比對於此酯化反應的影響，也在此做完整探討。此外，以擬均相數學模型來描述此含有固體觸媒之酯化反應行為。此數學模型更進一步的用來預測在轉化率為0.99的要求下，其初始醇酸莫耳比、反應溫度與反應時間分別是14.9、372.15K與9.5小時。這樣預測的結果與實際實驗值進行驗證，其相對誤差小於0.5％。

The aim of this study was to develop a microalgal cultivation system for intracellular lipid production and a solid catalytic process for biodiesel production. The motile intensity of microalgae, which was strongly related to specific growth rate, was used as the index for selecting a suitable cultivation medium. It was observed that the color of microalgae changed from green in the growth phase to yellow in the stationary phase during cultivation porcesses, mainly due to the changes of the biochemical composition in the microalgae. The color was analyzed by image analysis and an RGB (Red, Green, and Blue) model was developed to correlate the change in color to chlorophyll a and lipid contents of microalgae. The model was used to predict the chlorophyll a and lipid contents of microalgae and the experimental results were well correlated by the model (R2 > 0.94). This colorimetric method was more suitable in monitoring chlorophyll a and lipid contents in the microalgae as compared to the standard analytical methods.
The culture condition for lipid production differs from that of cell growth; it is possible to separate the growth and the production phases into two separate stages. The maximum lipid accumulation occurs under the nitrogen-limited growth. A two-stage cultivation strategy was developed. The microalgae was cultivated in a nitrogen-sufficient phase for cell growth and transferred into a nitrogen-deficient phase to induce the lipid production. The production of lipid obtained from the two-stage process was 2.82-fold higher than that from the traditional single stage cultivation.
Furthermore, a solid catalytic process for converting microalgal lipid into biodiesel was developed by using soybean oil as the source. Fatty acids obtained from enzymatic hydrolysis of soybean oil were esterified with methanol by using strong acidic cation-exchange resin as a heterogeneous catalyst. The kinetic study of esterification was carried out with different levels of catalyst loading, reaction temperature and molar ratio of methanol to fatty acids. The experimental data can be well-correlated by a second order kinetic equation. The kinetic model was used to predict the optimal operating conditions at a fatty acids conversion of 99%. The optimal operating conditions numerically calculated by the kinetic model were found to be reactants molar ratio of 14.9, reaction temperature of 99oC and reaction time of 9.5 h. The predicted results were verified with experimental results and the relative error was less than 0.5%.

總目錄
摘要    I
ABSTRACT    III
誌謝    V
總目錄    VI
圖目錄    IX
表目錄    XII
第一章 緒論    1
1-1 前言    1
1-2 藻類簡介    4
1-2-1 影響藻類培養之環境因子    5
1-2-1-1 光    5
1-2-1-2 碳源種類    7
1-2-1-3 溫度    8
1-2-1-4培養基組成    9
1-2-1-5 鹽度    9
1-2-1-6 酸鹼值    10
1-2-2 藻類培養系統介紹    10
1-2-2-1 開放式藻類培養系統    11
1-2-2-2 密閉式藻類培養系統    11
1-2-3 藻類養殖發展現況    14
1-3 研究動機與章節架構    16
第二章 實驗材料與方法    17
2-1 藻類培養系統    17
2-1-1 藻種與培養基組成    17
2-1-2 實驗設備    20
2-1-3藻類濃度之分析方法    21
2-1-4 光照強度測定    21
2-1-5 油脂含量之分析方法（傳統分析方法）    21
2-1-6 藻類運動特性分析方法    27
2-1-7 葉綠素含量之分析方法    28
2-1-8 藻類三原色之分析方法    29
2-1-9 光生化反應器    31
2-1-10 兩階段培養策略    34
2-2 生質柴油製程系統    35
2-2-1 實驗藥品    35
2-2-2 油脂水解反應    36
2-2-3 陽離子交換樹脂活化程序    36
2-2-4 酯化反應    37
2-2-5 脂肪酸轉化率分析    37
第三章 藻類之運動特性    39
3-1 前言    39
3-2 文獻回顧    41
3-3 結果與討論    46
3-3-1 不同培養基濃度之探討    46
3-3-2 不同氮源種類之探討    53
3-4 結語    59
第四章 影像分析技術於快速估計藻類葉綠素與油脂含量之應用    60
4-1 前言    60
4-2 文獻回顧    62
4-3 結果與討論    71
4-3-1 不同培養條件對於藻類生長、葉綠素a以及油脂含量之影響    71
4-3-1-1 光照強度之影響    72
4-3-1-2 氮源種類之影響    75
4-3-1-3 鹽度之影響    77
4-3-1-4 二氧化碳通氣比例之影響    79
4-3-2 影像分析技術應用於定量藻類顏色變化之研究    81
4-3-2-1 藻體顏色定性之探討    81
4-3-2-2 葉綠素與油脂預測方程式之建立    85
4-3-2-3 葉綠素a與油脂含量預測方程式應用於藻類在氣泡塔反應器培養過程中葉綠素a與油脂含量之監測    90
4-4 結語    94
第五章 兩階段藻類培養策略於油脂生產之應用    95
5-1 前言    95
5-2 文獻回顧    96
5-3 結果與討論    97
5-3-1 兩階段培養程序可行性研究    97
5-3-2 第兩階段培養程序之最適化操作條件探討    99
5-4 結語    104
第六章 固體酸觸媒於生質柴油製程之應用    105
6-1 前言    105
6-2 文獻回顧    107
6-3 反應速率式模型建立    111
6-4 結果與討論    114
6-4-1 觸媒用量對於脂肪酸酯化反應的影響    115
6-4-2 反應溫度對於脂肪酸酯化反應的影響    118
6-4-3 醇酸莫耳比對於脂肪酸酯化反應的影響    120
6-4-4 反應速率式參數之決定    122
6-4-4-1 反應速率常數    122
6-4-4-2 活化能    126
6-4-4-3 觸媒用量與正向反應和平衡反應的前指數因子線性關係之常數    128
6-4-5 模擬實驗    130
6-4-6 模型驗證、最適化條件操作與實驗驗證    131
6-5 結語    133
第七章 結論與未來展望    134
7-1 結論    134
7-2 未來展望    136
參考資料    138
符號說明    153
作者簡歷    155


圖目錄

圖1-1 利用油脂生產生化柴油之酯化反應式    4
圖1-2 光合效率與光照強度關係圖    6
圖2-1 利用血清瓶進行藻類保存示意圖    19
圖2-2 藻類特徵波長量測    25
圖2-3 藻類吸光值與細胞乾重檢量線    25
圖2-4 脂肪酸甲酯分析流程圖    26
圖2-5 影像設備裝置圖    27
圖2-6 三原色程式外觀操作圖    30
圖2-7 平板式光生化反應器裝置示意圖    32
圖2-8 氣泡塔反應器裝置示意圖    33
圖2-9 兩階段培養程序操作示意圖    34
圖3-1 在不同濃度培養基條件下，等鞭金藻之生長曲線    49
圖3-2 等鞭金藻培養於不同濃度培養基下之運動強度時間趨勢圖        50
圖3-3 等鞭金藻於不同濃度培養基下之運動強度與比生長速率對照圖    51
圖3-4 等鞭金藻於不同濃度培養基下之運動強度與其比生長速率線性關係圖    52
圖3-5 等鞭金藻培養於不同氮源種類下之生長曲線    55
圖3-6 等鞭金藻培養於不同氮源種類下之運動強度時間趨勢圖    56
圖4-1 光合作用機制簡要圖    62
圖4-2 葉綠素化學結構圖    63
圖4-3 多種類胡蘿蔔素化學結構：β-carotene(β胡蘿蔔素)；Zeaxanthin(玉米黃素)；Violaxanthin(堇黃素)；Lutein(葉黃素)； Neoxanthin(新黃素)。    64
圖4-4 藻藍素化學結構    64
圖4-5 在不同的光照強度下，藻類生長曲線以及葉綠素a含量之時間趨勢圖    74
圖4-6 在不同的氮源培養環境下，藻類生長曲線以及葉綠素a含量之時間趨勢圖    76
圖4-7 在不同鹽度的培養環境下，藻類生長曲線以及葉綠素a含量之時間趨勢圖    78
圖4-8 在二氧化碳通氣比例條件測試下，藻類生長曲線以及葉綠素a含量之時間趨勢圖    80
圖4-9 藻類在培養過程中顏色變化之情形    81
圖4-10 擬球藻三原色分析示意圖    83
圖4-11 三原色飽和度值、葉綠素a含量以及油脂含量之時間趨勢圖    84
圖4-12 擬球藻葉綠素a實際測定值與預測值對應圖    89
圖4-13 擬球藻油脂實際測定值與預測值對應圖    89
圖4-14 擬球藻培養於氣泡塔反應器中之顏色變化時間趨勢圖，以各原色的平均飽和度值來計量藻類所產生顏色之變化    92
圖4-15 擬球藻培養於氣泡塔反應器中之生長曲線    93
圖4-16 擬球藻培養於氣泡塔反應器中其葉綠素a與油脂累積含量之時間趨勢圖    93
圖5-1 擬球藻培養於單一階段培養程序與兩階段培養程序之細胞濃度與油脂含量時間趨勢圖    98
圖5-2 藻類初始培養濃度對於藻類油脂累積含量與油脂產率的影響    102
圖6-1 離子交換樹脂之物理特性與化學結構（A：強酸型陽離子交換樹脂；B：強鹼型陰離子交換樹脂）    110
圖6-2 脂肪酸轉化率在不同觸媒使用量實驗條件下之時間趨勢圖    116
圖6-3 固體觸媒使用量對於動力學參數之影響    117
圖6-4 脂肪酸轉化率在不同反應溫度實驗條件下之時間趨勢圖    119
圖6-5 脂肪酸轉化率在不同醇酸莫耳比實驗條件下之時間趨勢圖    121
圖6-6 在不同實驗操作條件下，正向反應速率常數之計算    124
圖6-7 正向反應速率、逆向反應速率以及平衡反應速率之Arrhenius-Van’t Hoff plot    127
圖6-8 平衡反應速率和正向反應速率的前指數因子與觸媒使用量線性關係圖    129
圖6-9 脂肪酸轉化率實際測定值與模擬值對應圖    130
圖6-10 反應溫度、初始醇酸莫耳比與反應時間在特定平衡轉化率下之關係圖    132

表目錄

表1-1 藻類培養之開放式系統與密閉式系統之比較    13
表1-2 台灣藻類生產量    15
表1-3 藻類產品的商業應用    15
表2-1  Walne培養基組成配方    18
表2-2 微量金屬溶液(trace metal solution)組成配方    19
表2-3 實驗儀器設備表    20
表2-4 酯化反應實驗中所使用之酸性離子交換樹脂的物理性質    35
表3-1 具有運動特性之藻類    45
表3-2 等鞭金藻培養於不同氮源種類之下的單因子變異數統計分析(ANOVA)：運動強度時間趨勢分析    57
表3-3 等鞭金藻培養於不同氮源種類下之最終油脂累積量    58
表4-1 在不同光照強度下，藻類在停滯期前後期之油脂含量    74
表4-2 在不同的氮源培養環境下，藻類在停滯期前後期之油脂含量    76
表4-3 在不同鹽度的培養環境下，藻類在停滯期前後期之油脂含量    78
表4-4 在二氧化碳通氣比例條件測試下，藻類在停滯期前後期之油脂含量    80
表4-5 藻類樣品三原色分析表，以圖4-10為例分別針對各原色的平均飽和度與變異係數進行分析    83
表4-6 實驗條件之設計以求得葉綠素a含量與油脂含量預測方程式之參數值    88
表5-1在不同初始培養濃度、鹽度與光照強度對於擬球藻於第二階段培養程序下，對於藻類最終生質重、油脂含量與油脂產率的影響    103
表6-1 脂肪酸酯化批次反應實驗參數設計    114
表6-2 在不同的實驗操作條件下，平衡常數與正向反應速率常數估計值    125

                                

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