Delineating the biosynthesis of gentamicin x2, the common precursor of the gentamicin C antibiotic complex.
Delineating the biosynthesis of gentamicin x2, the common precursor of the gentamicin C antibiotic complex.
复制标题
描述庆大霉素 X2（庆大霉素 C 抗生素复合物的常见前体）的生物合成
DOI：
10.1016/j.chembiol.2014.12.012
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发表时间：
2015-02-19
影响因子：
--
通讯作者：
Sun Y
中科院分区：
文献类型：
--
作者：
Huang C;Huang F;Moison E;Guo J;Jian X;Duan X;Deng Z;Leadlay PF;Sun Y
庆大霉素C复合物是世界范围内用于治疗严重革兰氏阴性细菌感染的氨基糖苷类抗生素的混合物。尽管具有临床重要性，但其生物合成途径的酶学仍然不清楚。我们在这里报告了从第一个形成的假三糖庆大霉素A2到庆大霉素X2的四个酶催化步骤的见解，庆大霉素X2是C复合物所有组分的最后一个常见中间体。我们使用了单个基因的靶向突变和体外部分途径的重构来证明A2的C-3″的二级醇功能首先转化为胺，由氧化还原酶GenD2和转氨酶GenS2的串联作用催化。然后，胺被s -腺苷-l-蛋氨酸（SAM）依赖的n-甲基转移酶GenN特异性甲基化，形成庆大霉素a。最后，C-4″上的c -甲基化，形成庆大霉素X2，由SAM依赖和钴胺依赖的自由基酶GenD1催化。四种酶将庆大霉素A2转化为庆大霉素X2 GenD1是一种罕见的钴胺素和自由基sam依赖的甲基转移酶。临床有价值的抗生素庆大霉素C复合物的所有五种成分都来自假三糖庆大霉素X2。Huang等人使用基因敲除和体外重组来鉴定催化这一关键中间体的最后步骤的四种途径酶。
Gentamicin C complex is a mixture of aminoglycoside antibiotics used worldwide to treat severe Gram-negative bacterial infections. Despite its clinical importance, the enzymology of its biosynthetic pathway has remained obscure. We report here insights into the four enzyme-catalyzed steps that lead from the first-formed pseudotrisaccharide gentamicin A2 to gentamicin X2, the last common intermediate for all components of the C complex. We have used both targeted mutations of individual genes and reconstitution of portions of the pathway in vitro to show that the secondary alcohol function at C-3″ of A2 is first converted to an amine, catalyzed by the tandem operation of oxidoreductase GenD2 and transaminase GenS2. The amine is then specifically methylated by the S-adenosyl-l-methionine (SAM)-dependent N-methyltransferase GenN to form gentamicin A. Finally, C-methylation at C-4″ to form gentamicin X2 is catalyzed by the radical SAM-dependent and cobalamin-dependent enzyme GenD1. Key portion of the pathway to a globally used antibiotic is defined Four enzymes convert gentamicin A2 into gentamicin X2 GenD1 is an unusual cobalamin- and radical SAM-dependent methyltransferase All five components of the clinically valuable antibiotic gentamicin C complex derive from the pseudotrisaccharide gentamicin X2. Huang et al. use gene knockouts and in vitro reconstitution to identify the four pathway enzymes catalyzing the final steps leading to this key intermediate.
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影响因子：
11.4
作者：
Linde, Liat;Kerem, Batsheva
通讯作者：
Kerem, Batsheva
影响因子：
4
作者：
Cuccarese, Michael F.;Singh, Amit;Amiji, Mansoor;O'Doherty, George A.
通讯作者：
O'Doherty, George A.
影响因子：
--
作者：
Guo J;Huang F;Huang C;Duan X;Jian X;Leeper F;Deng Z;Leadlay PF;Sun Y
通讯作者：
Sun Y
影响因子：
3.2
作者：
Huang, Fanglu;Spiteller, Dieter;Spencer, Jonathan B.
通讯作者：
Spencer, Jonathan B.
gntK 的鉴定，庆大霉素生物合成中紫胺 C-6â2 甲基化所需的基因
影响因子：
1.2
作者：
Hong, Wenrong;Yan, Lingbin
通讯作者：
Yan, Lingbin