此論文利用植入 2-酮戊二酸:鐵氧還蛋白氧化還原酶（2-oxoglutarate:ferredoxin oxidoreductase, OGOR）的大腸桿菌，將此菌株分別以絲氨酸（serine）和蘋果酸（malate）做三重複演化實驗，並比較其中之差異。所使用的生物反應器，運用LED燈的發射和接收來偵測光強度並搭配厭氧包以及無線傳輸模組達成在厭氧環境下傳送資料，測量出大腸桿菌生長過程中的光學密度（OD600）變化並計算生長速率（growth rate）。此演化實驗分成兩組進行，每組實驗分為兩階段，第一階段以絲氨酸（serine）為碳源進行演化，發現菌株演化後生長速率（growth rate）向上提升;第二階段以蘋果酸（malate）為碳源進行演化，並觀察兩組菌株、混營菌株的差異。

We report the Escherichia coli based on recombinant 2- oxoglutarate:ferredoxin oxidoreductase (OGOR),and this strain is maded with serine and malate respectively. Repeat the evolution experiment and compare the differences. The bioreactor we used uses the emission and reception of LED lights to detect the light intensity and is equipped with an anaerobic package and a wireless transmission module to transmit data in an anaerobic environment,and measure the optical density during the growth of Escherichia coli and calculate the growth rate. The evolution experiment was divided into two groups. Each group of experiments was divided into two stages. The first stage used serine as the carbon source for evolution. It was found that the growth rate of the strain increased after evolution. In the second stage, malate was used as the carbon source for evolution, and the differences of the two groups of strains and mixotrophic strains were observed.

致謝 I
中文摘要 II
英文摘要 III
目錄 IV
圖目錄 VII
表目錄 IX
一、 緒論 1
1-1研究動機 1
1-2文獻回顧 3
1-2-1固碳途徑 3
1-2-1-1卡爾文循環 4
1-2-1-2 還原性三羧酸循環 5
1-2-2利用菌株進行固碳反應 6
1-2-3 前人研究基礎 9
二、 實驗材料及方法 12
2-1實驗樣品和培養基配方 12
2-2菌株的製作和保存 14
2-3演化實驗準備和設計 15
2-3-1演化前準備 15
2-3-2 三重複演化實驗設計 15
2-4生物反應器裝置 16
2-4-1 硬體簡介 16
2-4-2 氣體條件及製造方式 18
2-4-3生物反應器設計 20
三、 實驗結果 22
3-1演化實驗步驟 22
3-2演化實驗細節 23
3-2-1 第一階段－以絲胺酸作為碳源 23
3-2-2第二階段－以蘋果酸作為碳源 27
3-3演化結果 30
3-3-1三重覆演化之菌株比較 30
3-3-2驗證能量途徑 33
四、 結論及討論 35
附錄 36
A-1演化實驗全部數據 36
A-2 ARDUINO 程式碼 38
A-3 大型生物反應器設計 47
A-4 太陽能電池產氫 49
A-5 中英對照表 51
參考文獻 53

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