Decreasing the ring size of a cyclic nonribosomal peptide antibiotic by in-frame module deletion in the biosynthetic genes
Decreasing the ring size of a cyclic nonribosomal peptide antibiotic by in-frame module deletion in the biosynthetic genes
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DOI：
10.1021/ja027276m
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发表时间：
2002-09-18
影响因子：
15
通讯作者：
Marahiel, MA
中科院分区：
文献类型：
--
作者：
Mootz, HD;Kessler, N;Marahiel, MA
许多具有治疗和生物技术重要性的天然产物是非核糖体合成的肽。这类化合物的结构特征是出现不寻常的氨基酸，主要是环状肽骨架，以及许多进一步的修饰，如酰化，杂环形成和糖基化。由于它们的结构复杂性，化学合成通常是一个没有吸引力的路线，这些分子。相比之下，生物合成基因的基因工程作为先导化合物的有用类似物的组合生物合成的潜在有力方法而出现。非核糖体肽合成酶（NRPS）在由多载体模型描述的硫模板过程中进行肽的连续多步组装和修饰。NRPS的模块化结构表明了通过交换催化亚基来重编程这些酶的简单方法。然而，许多报道的工程化尝试面临低产物产率或甚至无活性的杂合酶。使用一种新的方法来获得杂合NRPS，我们在这里表明，在NRPS装配线的整个模块的删除引起预测的肽抗生素变体的分泌与减少环的大小。此外，模块交换导致比先前研究中观察到的显著更高的产物产率。
Many natural products of therapeutical and biotechnological importance are nonribosomally synthesized peptides. Structural hallmarks of this class of compounds are the occurrence of unusual amino acids, mostly cyclic peptide backbones, and numerous further modifications such as acylation, heterocyclic ring formation, and glycosylation. Because of their structural complexity, chemical synthesis is usually an unattractive route to these molecules. In contrast, genetic engineering of the biosynthesis genes emerges as a potentially powerful approach to the combinatorial biosynthesis of useful analogues of the lead compounds. Nonribosomal peptide synthetases (NRPSs) carry out a sequential multistep assembly and modification of the peptides in a thiotemplate process described by the multiple carrier model. The modular architecture of NRPSs suggests straightforward methods for the reprogramming of these enzymes by exchange of catalytic subunits. However, many of the reported engineering attempts faced low product yields or even inactive hybrid enzymes. Using a new approach to obtain hybrid NRPSs, we show here that the deletion of an entire module in an NRPS assembly line caused the secretion of the predicted peptide antibiotic variant with a decreased ring size. Furthermore, a module exchange resulted in a significantly higher product yield than that observed in previous studies.