A Cell‐Free Platform Based on Nisin Biosynthesis for Discovering Novel Lanthipeptides and Guiding their Overproduction In Vivo
Abstract 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....
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2020-09-01
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| Series: | Advanced Science |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/advs.202001616 |
| _version_ | 1818968996741906432 |
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| author | Ran Liu Yuchen Zhang Guoqing Zhai Shuai Fu Yao Xia Ben Hu Xuan Cai Yan Zhang Yan Li Zixin Deng Tiangang Liu |
| author_facet | Ran Liu Yuchen Zhang Guoqing Zhai Shuai Fu Yao Xia Ben Hu Xuan Cai Yan Zhang Yan Li Zixin Deng Tiangang Liu |
| author_sort | Ran Liu |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-20T14:13:34Z |
| format | Article |
| id | doaj.art-8da37882c8a74da0a649b0e1d6bcb324 |
| institution | Directory Open Access Journal |
| issn | 2198-3844 |
| language | English |
| last_indexed | 2024-12-20T14:13:34Z |
| publishDate | 2020-09-01 |
| publisher | Wiley |
| record_format | Article |
| series | Advanced Science |
| spelling | doaj.art-8da37882c8a74da0a649b0e1d6bcb3242022-12-21T19:38:06ZengWileyAdvanced Science2198-38442020-09-01717n/an/a10.1002/advs.202001616A Cell‐Free Platform Based on Nisin Biosynthesis for Discovering Novel Lanthipeptides and Guiding their Overproduction In VivoRan Liu0Yuchen Zhang1Guoqing Zhai2Shuai Fu3Yao Xia4Ben Hu5Xuan Cai6Yan Zhang7Yan Li8Zixin Deng9Tiangang Liu10Key Laboratory of Combinatorial Biosynthesis and Drug Discovery Ministry of Education and Wuhan University School of Pharmaceutical Sciences Wuhan 430071 ChinaKey Laboratory of Combinatorial Biosynthesis and Drug Discovery Ministry of Education and Wuhan University School of Pharmaceutical Sciences Wuhan 430071 ChinaKey Laboratory of Combinatorial Biosynthesis and Drug Discovery Ministry of Education and Wuhan University School of Pharmaceutical Sciences Wuhan 430071 ChinaKey Laboratory of Combinatorial Biosynthesis and Drug Discovery Ministry of Education and Wuhan University School of Pharmaceutical Sciences Wuhan 430071 ChinaKey Laboratory of Combinatorial Biosynthesis and Drug Discovery Ministry of Education and Wuhan University School of Pharmaceutical Sciences Wuhan 430071 ChinaKey Laboratory of Combinatorial Biosynthesis and Drug Discovery Ministry of Education and Wuhan University School of Pharmaceutical Sciences Wuhan 430071 ChinaDepartment of Clinical Laboratory Renmin Hospital of Wuhan University Wuhan 430060 ChinaDepartment of Clinical Laboratory Renmin Hospital of Wuhan University Wuhan 430060 ChinaDepartment of Clinical Laboratory Renmin Hospital of Wuhan University Wuhan 430060 ChinaKey Laboratory of Combinatorial Biosynthesis and Drug Discovery Ministry of Education and Wuhan University School of Pharmaceutical Sciences Wuhan 430071 ChinaKey Laboratory of Combinatorial Biosynthesis and Drug Discovery Ministry of Education and Wuhan University School of Pharmaceutical Sciences Wuhan 430071 ChinaAbstract 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.https://doi.org/10.1002/advs.202001616cell‐free protein synthesisgenome mininglanthipeptideslanthipeptides overproductionlanthipeptides screening |
| spellingShingle | Ran Liu Yuchen Zhang Guoqing Zhai Shuai Fu Yao Xia Ben Hu Xuan Cai Yan Zhang Yan Li Zixin Deng Tiangang Liu A Cell‐Free Platform Based on Nisin Biosynthesis for Discovering Novel Lanthipeptides and Guiding their Overproduction In Vivo Advanced Science cell‐free protein synthesis genome mining lanthipeptides lanthipeptides overproduction lanthipeptides screening |
| title | A Cell‐Free Platform Based on Nisin Biosynthesis for Discovering Novel Lanthipeptides and Guiding their Overproduction In Vivo |
| title_full | A Cell‐Free Platform Based on Nisin Biosynthesis for Discovering Novel Lanthipeptides and Guiding their Overproduction In Vivo |
| title_fullStr | A Cell‐Free Platform Based on Nisin Biosynthesis for Discovering Novel Lanthipeptides and Guiding their Overproduction In Vivo |
| title_full_unstemmed | A Cell‐Free Platform Based on Nisin Biosynthesis for Discovering Novel Lanthipeptides and Guiding their Overproduction In Vivo |
| title_short | A Cell‐Free Platform Based on Nisin Biosynthesis for Discovering Novel Lanthipeptides and Guiding their Overproduction In Vivo |
| title_sort | cell free platform based on nisin biosynthesis for discovering novel lanthipeptides and guiding their overproduction in vivo |
| topic | cell‐free protein synthesis genome mining lanthipeptides lanthipeptides overproduction lanthipeptides screening |
| url | https://doi.org/10.1002/advs.202001616 |
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