A Cell-Free Platform Based on Nisin Biosynthesis for Discovering Novel Lanthipeptides and Guiding their Overproduction In Vivo.
A Cell-Free Platform Based on Nisin Biosynthesis for Discovering Novel Lanthipeptides and Guiding their Overproduction In Vivo.
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DOI：
10.1002/advs.202001616
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发表时间：
2020-09
期刊：
影响因子：
--
通讯作者：
Liu T
中科院分区：
文献类型：
--
作者：
Liu R;Zhang Y;Zhai G;Fu S;Xia Y;Hu B;Cai X;Zhang Y;Li Y;Deng Z;Liu T
羊毛硫肽具有广泛的治疗和工业应用。然而，由于许多是杀菌的，传统的体内平台在发现和大量生产新型羊毛硫肽的能力方面受到限制，因为细菌生物体通常是这些系统中的关键组分。在此，描述了无细胞蛋白质合成（CFPS）平台的开发，该平台使得能够在体内快速基因组挖掘、筛选和引导羊毛硫肽的过量生产。对于概念验证研究，选择I型羊毛硫肽，乳链菌肽。在美国国家生物技术信息中心（NCBI）数据库中的所有乳酸链球菌素类似物中，在一天内鉴定出四种具有抗菌活性的新羊毛硫肽。此外，CFPS平台与抗革兰氏阴性菌生长的筛选测定相结合，从而鉴定出有效的乳链菌肽突变体M5。发现乳链菌肽和乳链菌肽类似物的滴度在CFPS平台指导下得到改善。由于生物合成的相似性，CFPS平台广泛适用于其他羊毛硫肽，从而为羊毛硫肽的发现和过量生产提供了通用方法。描述了无细胞蛋白质合成（CFPS）平台的开发，用于快速基因组挖掘，筛选和指导体内羊毛硫肽的过量生产。CFPS平台的功能通过使用乳链菌肽（一种典型的羊毛硫肽）进行概念验证研究来证明。该CFPS平台为羊毛硫肽的发现和过量生产提供了通用方法。
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. The development of a cell‐free protein synthesis (CFPS) platform is described for rapid genome mining, screening, and the guiding overproduction of lanthipeptides in vivo. The functionality of the CFPS platform is demonstrated by using nisin (a typical lanthipeptide) for proof‐of‐concept studies. This CFPS platform provides a universal method for lanthipeptide discovery and overproduction.
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影响因子：
4
作者：
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