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研究生:傅怡菁
研究生(外文):Yi-Ching Fu
論文名稱:耐輻射奇異球菌在纖維素分解上的應用
論文名稱(外文):Application Deinococcus radiodurans on Cellulose Degradation
指導教授:劉仲康劉仲康引用關係
指導教授(外文):Jong-Kang Liu
學位類別:碩士
校院名稱:國立中山大學
系所名稱:生物科學系研究所
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:96
中文關鍵詞:紫外光纖維分解酵素纖維素游離性輻射細胞內粗萃物耐輻射奇異球菌轉殖菌株
外文關鍵詞:Escherichia colicelluloseradioactive wastemicroorganismsradiationThermotoga maritimaUVbioremediationDeinococcus radiodurans
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在受輻射污染之地區或一些輻射廢棄物中均含有大量之纖維素,造成廢棄物處理上的極大困擾。一般情形下都只能等輻射強度自然衰退達到背景值後才可進行一般的廢棄物之處理程序。由於傳統分解纖維素之微生物無法在輻射環境下生長,因此往往造成這些含輻射廢棄物內的有機污染物大量累積。一株耐輻射奇異球菌 (Deinococcus radiodurans) 對游離性輻射、紫外光及乾旱具有高度的抗性。因此可利用其特性來處理含輻射之廢棄物。經由本實驗得知,耐輻射奇異球菌在輻射環境下之生長與正常情形下差異不大。纖維分解酵素分解纖維素形成葡萄糖的實驗中發現,酵素活性並未受低劑量輻射影響但會受照射 UV 而影響酵素活性。但若另外加入 D. radiodurans 或其細胞內粗萃物即可降低 UV 對酵素活性的影響。本研究並已完成 Thermotoga maritima 纖維分解酵素之基因 celA 轉殖到 E. coli BL21 和 D. radiodurans 中,兩者皆有其酵素活性的表現。 Whole cell 照射 UV 對 celA 活性並無影響,但若將其細胞打破取其粗萃液持續照射 UV (強度 2.78 mw/cm2) ,則 E. coli 之 celA 活性會大幅降低 78 %,而 D. radiodurans 只降低些許約 18 %。此 D. radiodurans 轉殖菌株將可被利用在輻射污染的環境下分解廢棄物中的纖維素達到垃圾減量的目的。


There are large amount of cellulose accumulated in radioactive waste and radioactive pollution sites. It is difficult to clean up these cellulose. In general, waste treatment process can only proceed until the radiation decay to a safty level. Since most cellulolytic microorganisms could not survive in radioactive waste, the accumulation of cellulose in radioactive waste become a serious problem. Deinococcus radiodurans is highly resistant to radiation, UV light, and dryness. It is possible to use this bacterial strain in the bioremediation of radioactive waste. In this study, we found out that there was not much difference on the growth of this organism under radiation and UV light. Cellulose enzyme activity was inhibited by UV irradiation, but not by 32P radiation. The addition of D. radiodurans whole cells or its cell crude extracts could protect the cellulase from UV damage. We also successfully constructed two plasmids, that contained a cel A gene isolated from Thermotoga maritima. These two plasmids had been used to transform Escherichia coli BL21 and D. radiodurans. All transformed bacterial strains could express celA activity. The celA activities in these transformed D. radiodurans strains were not affect by UV irradiation. However, celA enzyme activity in the transformed E. coli was greatly inhibited by UV irradiation up to 78%. Hopefully these two transformed D. radiodurans bacterial strains can be applied to the bioremediation of radioactive waste.


目錄
中文摘要………………………………..……………………………...Ⅰ
英文摘要………………………………..……………………………...Ⅱ
壹、前言……………………………………………………………….1
貳、材料與方法………………………………………………………14
參、結果………………………………………………………………26
肆、討論………………………………………………………………37
伍、參考文獻…………………………………………………………45
陸、圖表………………………………………………………………53
柒、附錄………………………………………………………………86


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