Journal
ADVANCED SCIENCE
Volume 7, Issue 17, Pages -Publisher
WILEY
DOI: 10.1002/advs.202001616
Keywords
cell-free protein synthesis; genome mining; lanthipeptides; lanthipeptides overproduction; lanthipeptides screening
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Funding
- National Key R&D Program of China [2018YFA0900400]
- National Natural Science Foundation of China [31971341]
- Hubei Natural Science Fund Project [2017CFA054]
- J1 Biotech Co., Ltd.
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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.
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