A cell-free platform based on nisin biosynthesis for discovering novel lanthipeptides and guiding their overproduction in vivo
Posted 05 Sep 2019
bioRxiv DOI: 10.1101/757591 (published DOI: 10.1002/advs.202001616)
Posted 05 Sep 2019
Lanthipeptides have extensive therapeutic and industrial applications; however, since 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. We developed a cell-free protein synthesis (CFPS) platform that enables rapid genome mining, screening and guiding overproduction of lanthipeptides in vivo. For proof-of-concept studies, the type I lanthipeptide, nisin, was selected. Four novel lanthipeptides with anti-bacterial activity were identified among all nisin analogs in the NCBI database in a single day. Further, we coupled the CFPS platform with a screening assay for gram-negative bacterial growth and identified a potent nisin mutant, M5. The titer of nisin and nisin analogs significantly improved with CFPS platform guidance. Owing to the similarities in biosynthesis, our CFPS platform is broadly applicable to other lanthipeptides, provides a universal method for lanthipeptides discovery and overproduction.
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