Three enzymes involved in the N-methylation and incorporation of the pradimicin sugar moieties.
Three enzymes involved in the N-methylation and incorporation of the pradimicin sugar moieties.
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DOI：
10.1016/j.bmcl.2015.01.043
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发表时间：
2015-03-15
影响因子：
2.7
通讯作者：
Zhan, Jixun
中科院分区：
文献类型：
--
作者：
Napan, Kandy L.;Zhang, Shuwei;Anderson, Thomas;Takemoto, Jon Y.;Zhan, Jixun
Pradimicins是来自hibisca放线瘤P157-2的抗真菌和抗病毒天然产物。糖部分在这些化合物的生物活性中起着关键作用。在pradimicin生物合成基因簇中有两个糖基转移酶基因pdmS和pdmQ，它们被认为在pradimicin生物合成过程中负责糖部分的引入。在这项研究中，我们使用双交叉方法破坏了这两个基因。pdmS的破坏导致产生pradimicinone I， pradimicinone I是pradimicinin A的糖元，这证实了pdmS是负责第一个糖基化步骤的o-糖基转移酶，并将4',6'-二脱氧-4'-氨基-d -半乳糖或4',6'-二脱氧-4'-甲基氨基-d -半乳糖部分连接到5-OH。pdmQ的破坏导致pradimicin B的产生，表明该酶是第二个糖基转移酶，将d -木糖片段引入到第一个糖片段的3'- OH上。在pdmO之前插入整合质粒可能干扰了专用启动子，产生产生pradimicin C作为主要代谢物的突变体，这表明pdmO是特异性甲基化4',6'-二脱氧-4'-氨基-d -半乳糖片段的4'- NH2的酶。这些糖修饰和结合酶的功能表征有助于理解普拉地米星的生物合成途径。
Pradimicins are antifungal and antiviral natural products from Actinomadura hibisca P157-2. The sugar moieties play a critical role in the biological activities of these compounds. There are two glycosyltransferase genes in the pradimicin biosynthetic gene cluster, pdmS and pdmQ, which are putatively responsible for the introduction of the sugar moieties during pradimicin biosynthesis. In this study, we disrupted these two genes using a double crossover approach. Disruption of pdmS led to the production of pradimicinone I, the aglycon of pradimicin A, which confirmed that PdmS is the O-glycosyltransferase responsible for the first glycosylation step and attaching the 4',6'-dideoxy-4'-amino-D-galactose or 4',6'-dideoxy-4'-methylamino-D-galactose moiety to the 5-OH. Disruption of pdmQ resulted in the production of pradimicin B, indicating that this enzyme is the second glycosyltransferase that introduces the D-xylose moiety to the 3'- OH of the first sugar moiety. Insertion of an integrative plasmid before pdmO might have interfered with the dedicated promoter, yielding a mutant that produces pradimicin C as the major metabolite, which suggested that PdmO is the enzyme that specifically methylates the 4'- NH2 of the 4',6'-dideoxy-4'-amino-D-galactose moiety. Functional characterization of these sugar-decorating and –incorporating enzymes thus facilitates the understanding of the pradimicin biosynthetic pathway.
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影响因子：
2.9
作者：
Burgie, E. Sethe;Holden, Hazel M.
通讯作者：
Holden, Hazel M.
影响因子：
3.2
作者：
Napan, Kandy;Zhang, Shuwei;Morgan, Whitney;Anderson, Thomas;Takemoto, Jon Y.;Zhan, Jixun
通讯作者：
Zhan, Jixun
影响因子：
2.7
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3.2
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通讯作者：
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影响因子：
3.2
作者：
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通讯作者：
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