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Molecular Cloning of N-methylputrescine Oxidase from Tobacco

机译:烟草N-甲基腐胺氧化酶的分子克隆

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摘要

Nicotine biosynthesis in Nicotiana species requires an oxidative deamination of N-methylputrescine, catalyzed by N-methylputrescine oxidase (MPO). In a screen for tobacco genes that were down-regulated in a tobacco mutant with altered regulation of nicotine biosynthesis, we identified two homologous MPO cDNAs which encode diamine oxidases of a particular subclass. Tobacco MPO genes were expressed specifically in the root, and up-regulated by jasmonate treatment. Recombinant MPO protein expressed in Escherichia coli formed a homodimer and deaminated N-methylputrescine more efficiently than symmetrical diamines. These results indicate that MPO evolved from general diamine oxidases to function effectively in nicotine biosynthesis.
机译:烟草物种中的尼古丁生物合成需要N-甲基腐胺氧化酶(MPO)催化的N-甲基腐胺的氧化脱氨作用。在筛选烟草突变体中尼古丁生物合成调控改变的烟草基因中下调的筛选基因,我们鉴定了两个同源的MPO cDNA,它们编码特定亚类的二胺氧化酶。烟草MPO基因在根中特异性表达,并通过茉莉酸酯处理上调。在大肠杆菌中表达的重组MPO蛋白比同型二胺更有效地形成了均二聚体和脱氨基的N-甲基putrescine。这些结果表明,MPO由一般的二胺氧化酶演变为在尼古丁生物合成中有效发挥作用。

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