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    王丽慧, 丁庆豹, 欧伶, 许彦梅, 张春燕, 王玥. 酶法合成5-杂氮-2’-脱氧胞苷[J]. 华东理工大学学报(自然科学版), 2011, (3): 325-329.
    引用本文: 王丽慧, 丁庆豹, 欧伶, 许彦梅, 张春燕, 王玥. 酶法合成5-杂氮-2’-脱氧胞苷[J]. 华东理工大学学报(自然科学版), 2011, (3): 325-329.
    WANG Li-hui, DING Qing-bao, OU Ling, XU Yan-mei, ZHANG Chun-yan, WANG Yue. Enzymatic Synthesis of 5-Aza-2’-Deoxycytidine[J]. Journal of East China University of Science and Technology, 2011, (3): 325-329.
    Citation: WANG Li-hui, DING Qing-bao, OU Ling, XU Yan-mei, ZHANG Chun-yan, WANG Yue. Enzymatic Synthesis of 5-Aza-2’-Deoxycytidine[J]. Journal of East China University of Science and Technology, 2011, (3): 325-329.

    酶法合成5-杂氮-2’-脱氧胞苷

    Enzymatic Synthesis of 5-Aza-2’-Deoxycytidine

    • 摘要: 将瑞士乳杆菌(Lactobacillus helveticus)中的N-脱氧核糖转移酶II的基因(ntd)克隆到大肠杆菌BL21(DE3)中,构建原核表达系统,成功地得到一株大量表达N-脱氧核糖转移酶II的菌株PND。以异丙基硫代半乳糖苷(IPTG)作为诱导剂,该重组菌可大量表达N-脱氧核糖转移酶II。在此酶作用下,以5-杂氮胞嘧啶和脱氧胸苷为底物,生成胸腺嘧啶和目标产物5-杂氮-2’-脱氧胞苷。最佳反应条件为:底物浓度均为2 mmol/L, 20 mmol/LTris-HCl缓冲液(pH为7.1),

       

      Abstract: The enzyme (N-deoxyribosyltransferase-II) gene (ntd) in Lactobacillus helveticus was cloned into E.coli BL21(DE3)to recombine a prokaryotic expression bacteria PND. The bacteria PND expressed large amounts of N-deoxyribosyltransfer ase-II with IPTG as an inducement. The 5-aza-2’-deoxycytidine and thymine were synthesied at the substrate of 5-azacytosine and deoxythymidine by the enzyme of N-deoxyribosyltrans-ferase-Ⅱ. The optimum condition was 2 mmol/L of subtrates, 20 mmol/L of Tris-HCl (pH 7.1), 200 μL of enzyme, 2 mL of reaction volume, and 2 h of reaction time at 55 ℃. Conversion of substrate reached 61% at the optimum condition.

       

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