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Student Number 89224002
Author Yin-Chieh Kuo(郭銀杰)
Author's Email Address ffjkuo@yahoo.com.tw
Statistics This thesis had been viewed 2760 times. Download 10 times.
Department Life Science
Year 2001
Semester 2
Degree Master
Type of Document Master's Thesis
Language zh-TW.Big5 Chinese
Title The purification of Triton X-100 oxygenase
Date of Defense 2002-07-02
Page Count 63
Keyword
  • Triton X-100
  • Abstract Triton X-100, octylphenol polyethoxylates, belongs to nonionic surfactant. Alkyl phenol polyethoxylates are used in numberous commercial and industrical products including detergent, dispersants and emalsifiers. Large quantity of surfactants released to the enviroment result in the influence of the fate of organic compound in soils. The degraded products, alkyl phenol derivative, were demonstrated environmental hormone effect. Our previous result suguessed that the degradation of this compound by a bacteria isolated by our lab is by a sequential cleavage of the ethoxylate chain. The enzyme activity involved in the scission of ethoxylate chain was monitored by oxygen uptake rate. The ether cleavage enzyme was demonstrated as a NADPH-dependent oxygenase. The purification of the bound enzyme was carried out by DEAE-Sepharose, ammonium sulfate precipitate, Phenyl-Sepharose, MonoQ. The molecular weight of purified enzyme was 44 kD,analyzed by SDS-PAGE. The N-terminal amino acid was blocked. Therefore a trypsin-digested peptide were sequenced by Q-ToF MS. The two peptides sequences matched to an elongation factor Tu (EF-Tu). EF-Tu has been studied for many year in translation. It promotes aminoacyl-tRNA into the ribosomal A site from an EF-Tu· GTP· aminoacyl-tRNA complex with hydrolysis of GTP. The 5-10 % of total cell protein is EF-Tu and the excess over the other proteins may have other function (Kurland, et al., 1995). The recent studies has been shown that this protein may function as chaperon and thioredoxin. It plays a role in the protection of other protein from heat and osmotic stress. Whether this protein show an enzymatic activity in the oxygenation of oxygen into Triton X-100 needs to be confirmed.
    Table of Content 目錄………………………………………………………………….........I
    表目錄……………………………………………………………….........Ⅱ
    圖目錄….………………………………………………………….….......Ⅲ
    壹、緒論………………………………………………………….…........1
     一、界面活性劑……………………...............................1
     二、界面活性劑對環境的影響………………………………...........2
     三、界面活性劑之分解….…………………………………............2
     四、環境荷爾蒙……………………….............................3
    五、蛋白體學研究...............................................4
    六、研究背景與大綱.............................................5
    貳、材料與方法…………………………………………………….........8
    一、細菌培養與培基…………………………………….................8
     二、細菌形態…………………………………….....................9
     三、酵素純化……………………………….........................9
     四、胺基酸定序……………………………….......................12
     五、二維電泳…………………………………….....................13
    參、結果…………………………………………………………….........20
    一、細菌形態…………………………...............................20
    二、酵素純化與序……………………...............................20
    三、二維電泳分析…………………………...........................22
    肆、討論……………………………………………………….............23
    一、菌種形態...................................................23
    二、酵素純化……………………….................................23
    伍、建議………………………………………………….................29
    陸、參考文獻….................................................30
    表…………………………………………………………………….........36
    圖…………………………………………………………………….........40
    附錄...........................................................56
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    Advisor
  • Shir-Ly Huang(黃雪莉)
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    Date of Submission 2002-07-15

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