Gene inactivation study of gntE reveals its role in the first step of pseudotrisaccharide modifications in gentamicin biosynthesis
Gene inactivation study of gntE reveals its role in the first step of pseudotrisaccharide modifications in gentamicin biosynthesis
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DOI：
10.1016/j.bbrc.2008.05.133
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发表时间：
2008-08-08
影响因子：
3.1
通讯作者：
Kwon, Hyung-Jin
中科院分区：
文献类型：
--
作者：
Kim, Jin-Yong;Suh, Joo-Won;Kwon, Hyung-Jin
对gntE（棘孢小单孢菌中庆大霉素生物合成基因簇的成员）进行基因失活研究。计算机辅助同源性分析预测GntE中的甲基转移酶相关钴胺素结合结构域和自由基S-腺苷甲硫氨酸结构域。在其他氨基糖苷类生物合成基因簇中没有发现gntE同源物。在两种M中均实现gntE的失活。棘孢菌ATCC 15835和庆大霉素高产菌株GMC 106。高效液相色谱分析，再加上质谱，显示gntE突变体积累庆大霉素A2及其衍生物与安装在葡糖胺部分的甲基。该结果证实GntE参与庆大霉素生物合成中假三糖修饰的第一步，尽管这种不寻常的氧化还原酶/甲基转移酶候选物的催化性质尚未解决。目前的基因失活研究还表明，靶向基因工程可以应用于生产特定的庆大霉素结构和潜在的新的庆大霉素衍生物在M。棘孢菌属。(c)2008年爱思唯尔公司All rights reserved.
A gene inactivation study was performed on gntE, a member of the gentamicin biosynthetic gene cluster in Micromonospora echinospora. Computer-aided homology analysis predicts a methyltransferase-related cobalamin-binding domain and a radical S-adenosylmethionine domain in GntE. It is also found that there is no gntE homolog within other aminoglycoside biosynthetic gene clusters. Inactivation of gntE was achieved in both M. echinospora ATCC 15835 and a gentamicin high-producer GMC106. High-performance liquid chromatographic analysis, coupled with mass spectrometry, revealed that gntE mutants accumulated gentamicin A2 and its derivative with a methyl group installed on the glucoamine moiety. This result substantiated that GntE participates in the first step of pseudotrisaccharide modifications in gentamicin biosynthesis, though the catalytic nature of this unusual oxidoreductase/methyltransferase candidate is not resolved. The present gene inactivation study also demonstrates that targeted genetic engineering can be applied to produce specific gentamicin structures and potentially new gentamicin derivatives in M. echinospora. (c) 2008 Elsevier Inc. All rights reserved.