臺灣博碩士論文加值系統

摘要
耐輻射奇異球菌(Deinococcus radiodurans)以胰化蛋白─葡萄糖─酵母萃取液(tryptone-glucose-yeast extract, 簡稱TGY培養基) 培養時，具有高度輻射抗性。此時細胞會利用磷酸五碳糖代謝途徑(pentose phosphate pathway, PPP) 緩慢代謝葡萄糖，而不會表現糖解作用(glycolytic Embden-Meyerhof pathway, EMP) 的活性。但是培養至靜止期時加入10 µM二價錳離子之後，可誘發二次生長(Mn-CD效應) 以及EMP pathway，代謝葡萄糖的比例大約是EMP:PPP=6:1，而且輻射抗性下降。分析與DNA修補相關的代謝物，發現加入錳離子之後的菌體內的IMP (inosine monophosphate)、UMP (uridine monophosphate) 以及NAD (nicotine adenine dinucleotide) 濃度明顯降低。如果更換培養基內碳源為果糖(TFY培養基) 或醋酸鈉(TAY培養基)，或者是只有用TY培養基培養，代謝物濃度變化仍然是加錳離子之後較低。除此之外，我們也發現以TGY與TFY培養時PPP代謝物濃度較高，以TAY與TY培養時較低。此發現證實葡萄糖與果糖係經由PPP路徑來進行代謝。所有培養的細胞經過UV照射以後IMP、UMP與NAD濃度均明顯下降，顯示這些代謝物可能被使用於DNA之修補。

Abstract
Deinococcus radiodurans is highly resistant to radiation when it grown in tryptone-glucose-yeast extraxt (TGY) medium. It oxidized glucose slowly mainly by the pentose phosphate pathway (PPP) and showed little glycolytic Embden-Meyerhof pathway (EMP) activity. The addition of 10 µM Mn(II) into the stationary phase cultures, could induced new round of cell division (Mn-CD effect) and the EMP activity. Glucose metabolized by Mn-CD cells at a EMP:PPP=6:1 ratio. In analyzing the metabolites for DNA repair, we found that after the addition of Mn(II) , the concentrations of PPP metabolites such as insione monophosphate (IMP)、uridine monophosphate (UMP) and NAD (nicotine adenine dinucleotide) were greatly reduced. This event is also occurred when replacing the glucose by fructose, sodium acetate, or removing glucose from the TGY culture medium. Besides, we also found that the TGY and TFY grown cells contained more PPP metabolites than those of TAY and TY cells. This finding suggested that glucose and fructose were metabolized by the PPP pathway in D. radiodurans. Finally, the concentrations of IMP、UMP and NAD in the cells were greatly decreased after UV irradiation. This indicated that these metabolites were probably employed to repair the DNA damage causing by UV irradiation.

目錄
中文摘要………………………………………………………………I
英文摘要……………………………………………………………II
圖目錄………………………………………………………………..III
前言……………………………………………………………………1
材料與方法…………………………………………………………13
結果……………………………………………………………………19
討論……………………………………………………………………26
參考資料…………………………….……………………………..34
圖………………………………………………………………………42
附圖……………………………………………………………………64

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