Lanthipeptides have extensive therapeutic and industrial applications. However, because many are bactericidal, traditional in vivo platforms are limited in their capacity to discover and mass produce novel lanthipeptides as bacterial organisms are often critical components in these systems. Herein, the development of a cell‐free protein synthesis (CFPS) platform that enables rapid genome mining, screening, and guided overproduction of lanthipeptides in vivo is described. For proof‐of‐concept studies, a type I lanthipeptide, nisin, is selected. Four novel lanthipeptides with antibacterial activity are identified among all nisin analogs in the National Center for Biotechnology Information (NCBI) database in a single day. Further, the CFPS platform is coupled with a screening assay for anti‐gram‐negative bacteria growth, resulting in the identification of a potent nisin mutant, M5. The titers of nisin and the nisin analog are found to be improved with CFPS platform guidance. Owing to the similarities in biosynthesis, the CFPS platform is broadly applicable to other lanthipeptides, thereby providing a universal method for lanthipeptide discovery and overproduction.

中文翻译：

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基于乳链菌肽生物合成的无细胞平台，用于发现新型多肽和指导其体内过量生产

脂肽具有广泛的治疗和工业应用。但是，由于许多细菌具有杀菌作用，因此传统的体内平台在发现和大规模生产新型多肽肽方面的能力受到限制，因为细菌生物通常是这些系统中的关键组成部分。本文描述了无细胞蛋白质合成（CFPS）平台的开发，该平台可在体内快速进行基因组挖掘，筛选和指导超量生产多肽。为了进行概念验证研究，选择了I型乳肽乳链菌肽。一天之内，在国家生物技术信息中心（NCBI）数据库中的所有乳链菌肽类似物中，鉴定出四种具有抗菌活性的新型脂肽。此外，CFPS平台还结合了用于抗革兰氏阴性细菌生长的筛选测定，结果鉴定出有效的乳链菌肽突变体M5。发现使用CFPS平台指南可改善乳链菌肽和乳链菌肽类似物的效价。由于生物合成方面的相似性，CFPS平台可广泛应用于其他多肽，从而为发现和过量生产多肽提供了一种通用方法。