倪现朴， 张宏宇， 夏焕章^*

（  沈阳药科大学生命科学与生物制药，沈阳 110016；  ）

  职称论文发表摘要： 庆大霉素是临床常用的氨基糖苷类抗生素，庆大霉素结构中存在四个甲基。甲基化对庆大霉素生物活性和毒性有重要影响。而庆大霉素生物合成中4''-甲基转移酶基因并未研究清楚。为阐明4''-甲基转移酶基因，对推测的甲基转移酶基因genN进行基因阻断与回补实验。构建、筛选获得genN阻断菌株，对该菌株发酵培养后发酵产物进行分离纯化。阻断菌株不再产生庆大霉素，而产生4''-脱甲基庆大霉素，将完整的genN基因回补到阻断菌株后，回补菌株又能够产生庆大霉素，证明genN负责庆大霉素生物合成中4''-甲基转移。

关键词： 微生物药学；棘孢小单孢菌；甲基转移酶；4''-脱甲基庆大霉素

NI Xianpu^1,， ZHANG Hongyu^2,， XIA Huanzhang^2,*

（  1、School of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University, Shenyang, Liaoning, 110016 ；       2、School of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University, Shenyang, Liaoning, 110016；  ）

Abstract： Gentamicin is widely used in clinic, which contains four major components differing from methyl groups. The methyl groups have a significant impact on antibiotic activity and toxicity of gentamicin. However, gene encoding methyltransferse that acts on C-4'' of gentamicin remains obscure. Sequence analysis has predicted that genN was a methyltransferase in gentamicin biosynthetic pathway. To study the function of genN, it was disrupted in the gentamicin producers M. echinospora. In genN disrupting strains, the production of gentamicin C complex was abolished and 4''-demethyl-gentamicin C complex was obtained. The restoration of gentamicin by self complementation suggested that there was no polar effect in the mutants. These gene inactivation and complementation studies suggested that GenN is C-methyltransferase that acts on C-4'' in gentamicin biosynthesis.
Keywords： microbiological pharmacy; Micromonospora echinospora; methyltransferase; 4''-demethyl-gentamicin

 

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