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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Cloning and characterization of the biosynthetic gene cluster for kutznerides
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Cloning and characterization of the biosynthetic gene cluster for kutznerides

机译:苦瓜生物合成基因簇的克隆与鉴定

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摘要

Kutznerides, actinomycete-derived cyclic depsipetides, consist of six nonproteinogenic residues, including a highly oxygenated tricyclic hexahydropyrroloindole, a chlorinated piperazic acid, 2-(1-methylcyclopropyl)-glycine, a β-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 f unctionalized 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.
机译:Kutznerides是由放线菌衍生的环状十足肽,由六个非蛋白质残基组成,包括高度氧化的三环六氢吡咯并吲哚,氯化哌嗪酸,2-(1-甲基环丙基)-甘氨酸,β-支链羟基酸和3-羟基谷氨酸,尚未阐明其生物合成逻辑。本文中,我们描述了库兹那利德家族的生物合成基因簇,通过简并引物PCR鉴定了卤化酶,这些卤化酶被假定与这些异常单体的生物合成有关。 56kb的基因簇编码一系列分布在三种蛋白质和各种定制酶(包括单核非血红素铁和两种黄素依赖性卤化酶)上的一系列六个非核糖体肽合成酶(NRPS)模块,以及一系列氧气转移催化剂。 NRPS基因的序列和组织支持将异常的单体单元掺入到库兹涅利德的稠密功能化支架中。我们的工作提供了对这类有趣的化合物形成的见解,并为阐明其生物合成的时机和机理提供了基础。

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