Non-ribosomal peptide synthetases (NRPSs) are the origin of a wide range of natural products, including many clinically used drugs. Engineering of these often giant biosynthetic machineries to produce novel non-ribosomal peptides (NRPs) at high titre is an ongoing challenge. Here we describe a strategy to functionally combine NRPS fragments of Gram-negative and -positive origin, synthesising novel peptides at titres up to 290 mg l^-1. Extending from the recently introduced definition of eXchange Units (XUs), we inserted synthetic zippers (SZs) to split single protein NRPSs into up to three independently expressed and translated polypeptide chains. These synthetic type of NRPS (type S) enables easier access to engineering, overcomes cloning limitations, and provides a simple and rapid approach to building peptide libraries via the combination of different NRPS subunits.

中文翻译：

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合成拉链作为非核糖体肽合成酶工程设计的辅助工具

非核糖体肽合成酶（NRPS）是多种天然产品的起源，包括许多临床使用的药物。对这些通常巨大的生物合成机器进行工程设计以产生高滴度的新型非核糖体肽（NRP）是一项持续的挑战。在这里，我们描述了一种策略，功能上结合革兰氏阴性和阳性来源的NRPS片段，在滴定度高达290 mg l ^-1时合成新肽。扩展自最近引入的e X change U的定义尼特（XU），我们插入了合成拉链（SZ），以将单个蛋白质NRPS分为多达三个独立表达和翻译的多肽链。这些合成类型的NRPS（S型）使工程更容易获得，克服了克隆限制，并通过不同NRPS亚基的组合提供了一种简单快速的方法来构建肽库。