Radical SAM-dependent ether crosslink in daropeptide biosynthesis.
Radical SAM-dependent ether crosslink in daropeptide biosynthesis.
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DOI：
10.1038/s41467-022-30084-2
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发表时间：
2022-04-29
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
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作者：
Darobactin是一种核糖体合成和后修饰的肽（RiPP），其对多种革兰氏阴性菌具有有效的活性。Darobactin具有高度独特的双环骨架，由两个Trp残基之间的醚交联和Lys与Trp之间的C-C交联组成。在这里，我们报告在体内和体外活性的darobactin合酶DarE。我们表明DarE是一种自由基S-腺苷甲硫氨酸（rSAM）酶，并且仅负责形成darobactin的双环支架。DarE在体外主要产生醚交联产物，当在H218 O中进行测定时，观察到明显的18 O掺入到醚交联产物中。这些观察结果表明，在darobactin生物合成中存在rSAM依赖性过程，涉及水分子的极不寻常的氧插入步骤以及随后的O-H和C-H活化。基因组挖掘分析表明，darobactin样生物合成基因簇的多样性，其中一个亚支可能编码单环产物，只有一个醚键。我们建议将Daropeptide命名为DarE酶产生的含醚RIPP家族。Darobactin是一种核糖体合成和翻译后修饰的肽，具有独特的支架和针对革兰氏阴性细菌的强效活性。在这里，作者确定了darobactin合酶DarE负责双环支架的形成，并为这类不断增长的酶提出了daropeptide的名称。
Darobactin is a ribosomally synthesized and post-translationally modified peptide (RiPP), which possesses potent activity against various Gram-negative bacteria. Darobactin features a highly unique bicyclic scaffold, consisting of an ether crosslink between two Trp residues and a C–C crosslink between a Lys and a Trp. Here we report in vivo and in vitro activity of darobactin synthase DarE. We show DarE is a radical S-adenosylmethionine (rSAM) enzyme and is solely responsible for forming the bicyclic scaffold of darobactin. DarE mainly produced the ether-crosslinked product in vitro, and when the assay was performed in H218O, apparent 18O incorporation was observed into the ether-crosslinked product. These observations suggested an rSAM-dependent process in darobactin biosynthesis, involving a highly unusual oxygen insertion step from a water molecule and subsequent O–H and C–H activations. Genome mining analysis demonstrates the diversity of darobactin-like biosynthetic gene clusters, a subclade of which likely encode monocyclic products with only an ether linkage. We propose the name daropeptide for this growing family of ether-containing RiPPs produced by DarE enzymes. Darobactin is a ribosomally synthesized and post-translationally modified peptide featuring a unique scaffold and potent activity against Gram-negative bacteria. Here, the authors identify darobactin synthase DarE as responsible for the bicyclic scaffold formation and propose the name daropeptide for this growing class of enzymes.
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影响因子：
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影响因子：
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作者：
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DOI：
10.1074/jbc.ra120.015371
发表时间：
2020-12-04
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Balty C;Guillot A;Fradale L;Brewee C;Lefranc B;Herrero C;Sandström C;Leprince J;Berteau O;Benjdia A
通讯作者：
Benjdia A