酶法合成5-杂氮-2'-脱氧胞苷

Li-hui WANG,Qing-bao DING,Ling OU,Yan-mei XU,Chun-yan ZHANG,Yue WANG

Huadong Ligong Daxue Xuebao/Journal of East China University of Science and Technology（2011）

Cited 24|Views16

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Abstract

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),200 μL酶,最终反应体积2 mL,55℃反应2 h,转化率可达61%.

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Key words

5-aza-2'-CdR,N-deoxyribosyltransferase-II,Synthesis

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