Modification and de novo design of non-ribosomal peptide synthetases using specific assembly points within condensation domains
Modification and de novo design of non-ribosomal peptide synthetases using specific assembly points within condensation domains
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DOI：
10.1038/s41557-019-0276-z
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发表时间：
2019-07-01
期刊：
影响因子：
21.8
通讯作者：
Bode, Helge B.
中科院分区：
文献类型：
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作者：
Bozhueyuek, Kenan A. J.;Linck, Annabell;Bode, Helge B.
非核糖体肽合成酶（NRPSs）是一种巨大的酶机器，它以流水线的方式激活氨基酸。由于NRPSs不局限于20种蛋白质原氨基酸的结合，它们的有效操作将使微生物能够生产各种肽；然而，重编程NRPSs以促进新肽的产生的结构要求尚不清楚。在这里，我们描述了在NRPSs的缩合域内的一个新的融合点，导致了交换单元缩合域（XUC）概念的发展，这使得高效生产肽，甚至含有非天然氨基酸，产量高达280 mg l（-1）。这使得产生更特异的NRPSs，减少了不需要的肽衍生物的数量，同时也产生了肽库。因此，XUC可能适用于未来肽生产的优化和生物活性肽衍生物的鉴定，用于制药和其他应用。
Non-ribosomal peptide synthetases (NRPSs) are giant enzyme machines that activate amino acids in an assembly line fashion. As NRPSs are not restricted to the incorporation of the 20 proteinogenic amino acids, their efficient manipulation would enable microbial production of a diverse range of peptides; however, the structural requirements for reprogramming NRPSs to facilitate the production of new peptides are not clear. Here we describe a new fusion point inside the condensation domains of NRPSs that results in the development of the exchange unit condensation domain (XUC) concept, which enables the efficient production of peptides, even containing non-natural amino acids, in yields up to 280 mg l(-1). This allows the generation of more specific NRPSs, reducing the number of unwanted peptide derivatives, but also the generation of peptide libraries. The XUC might therefore be suitable for the future optimization of peptide production and the identification of bioactive peptide derivatives for pharmaceutical and other applications.