Cloning and characterization of the biosynthetic gene cluster for kutznerides
Cloning and characterization of the biosynthetic gene cluster for kutznerides
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DOI：
10.1073/pnas.0708242104
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发表时间：
2007-10-16
影响因子：
11.1
通讯作者：
Walsh, Christopher T.
中科院分区：
文献类型：
--
作者：
Fujimori, Danica Galonic;Hrvatin, Sinisa;Walsh, Christopher T.
Kutznerides是一种由放线菌衍生的环状去脂环，由六个非蛋白生成残基组成，包括一个高氧三环六氢吡咯吲哚、一个氯代哌嗪酸、2-(1-甲基环丙基)-甘氨酸、一个β-支链-羟基酸和3-羟基谷氨酸，其生物合成逻辑尚未阐明。在这里，我们描述了kutzneride家族的生物合成基因簇，通过简并引物PCR鉴定，卤化酶被认为参与了这些不寻常单体的生物合成。56kb的基因簇编码一系列六个非核糖体多肽合成酶(NRPS)模块，分布在三种蛋白质和各种剪裁酶上，包括单核非血红素铁和两个黄素依赖的卤化酶，以及一系列氧转移催化剂。NRPS基因的序列和组织支持将不寻常的单体单位掺入到库茨内里德的密集功能化支架中。我们的工作为这类有趣的化合物的形成提供了洞察力，并为阐明它们的生物合成的时间和机制提供了基础。
Kutznerides, actinomycete-derived cyclic depsipeticles, consist of six nonproteinogenic residues, including a highly oxygenated tricyclic hexahydropyrroloindole, a chlorinated piperazic acid, 2-(1methylcyclopropyl)-glycine, a beta-branched-hydroxy acid, and 3-hydroxy glutamic acid, for which biosynthetic logic has not been elucidated. Herein we describe the biosynthetic gene cluster for the kutzneride family, identified by degenerate primer PCR for halogenating enzymes postulated to be involved in biosyntheses of these unusual monomers. The 56-kb gene cluster encodes a series of six nonribosomal peptide synthetase (NRPS) modules distributed over three proteins and a variety of tailoring enzymes, including both mononuclear nonheme iron and two flavin-dependent halogenases, and an array of oxygen transfer catalysts. The sequence and organization of NRPS genes support incorporation of the unusual monomer units into the densely functionalized scaffold of kutznerides. Our work provides insight into the formation of this intriguing class of compounds and provides a foundation for elucidating the timing and mechanisms of their biosynthesis.