文化大學機構典藏 CCUR:Item 987654321/25209
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    請使用永久網址來引用或連結此文件: https://irlib.pccu.edu.tw/handle/987654321/25209


    題名: 牛分枝結核桿菌之支鏈胺基酸轉胺酶的定性分析 及其抑制劑之篩選
    Characterization of Mycobacterium bovis Branched-Chain Amino Acid Aminotransferase and Screening for Its Inhibitors
    作者: 阮氏梅香
    貢獻者: 生科所
    關鍵詞: 牛分枝結核桿菌
    Mycobacterium bovis
    日期: 2013-07
    上傳時間: 2013-09-12 15:43:57 (UTC+8)
    摘要: Tuberculosis mostly caused by Mycobacterium tuberculosis or Mycobacterium bovis is one of the most dangerous infectious diseases; therefore, it requires searching and discovering new medicines against mycobacteria. Polyamine synthesis and its associated methionine (Met) regeneration pathway have been found to be good drug targets in a variety of microorganisms. The conversion of ketomethiobutyrate (KMTB) to methionine is the final step in this pathway catalyzed by branched-chain amino acid aminotransferases (BCATs).
    In this study, the gene encoding for M. bovis BCAT (MbBCAT) was cloned, over-expressed, purified and characterized. Results showed that MbBCAT have optimal temperature at 40-45℃ and optimal pH at near pH 8.5. The kinetic parameters Km, kcat of MbBCAT were 0.12 M, 30 s-1 and 10.34M and 50 s-1 for ketoisocaproate and Glutamte, respectively. In the anlaysis of substrate specificity analysis, KIC and KMV were found to be the best amino-acceptor substrate when glutamte was used as amino-donor. For screening inhibitors against MbBCAT, molecular docking software (Autodock version 1.4.2) was used to evaluate the free energy status after the target enzyme MbBCAT was docked with ligands from chemical banks. Approximately 100,000 compounds were screened in silico, and 10 compounds with less free energy (less than -9) were selected as a possible drug candidate. Among these candidates, Congo Red and Sudan Black were found to possess inhibitory effect on MbBCAT with Ki about 6.5 M and 15.3 M, respectively. The antimycobacterial effects against M. bovis need to cooperate with other labs to further examination for evaluating its promissing for tuberculosis treatment.
    顯示於類別:[生物科技研究所 ] 博碩士論文

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