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Module and individual domain deletions of NRPS to produce plipastatin derivatives in Bacillus subtilis
BACKGROUND: Plipastatin, an antifungal lipopeptide, is synthesized by a non-ribosomal peptide synthetase (NRPS) in Bacillus subtilis. However, little information is available on the combinatorial biosynthesis strategies applied in plipastatin biosynthetic pathway. In this study, we applied module or...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5984369/ https://www.ncbi.nlm.nih.gov/pubmed/29855381 http://dx.doi.org/10.1186/s12934-018-0929-4 |
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author | Gao, Ling Guo, Jianping Fan, Yun Ma, Zhi Lu, Zhaoxin Zhang, Chong Zhao, Haizhen Bie, Xiaomei |
author_facet | Gao, Ling Guo, Jianping Fan, Yun Ma, Zhi Lu, Zhaoxin Zhang, Chong Zhao, Haizhen Bie, Xiaomei |
author_sort | Gao, Ling |
collection | PubMed |
description | BACKGROUND: Plipastatin, an antifungal lipopeptide, is synthesized by a non-ribosomal peptide synthetase (NRPS) in Bacillus subtilis. However, little information is available on the combinatorial biosynthesis strategies applied in plipastatin biosynthetic pathway. In this study, we applied module or individual domain deletion strategies to engineer the plipastatin biosynthetic pathway, and investigated the effect of deletions on the plipastatin assembly line, as well as revealed the synthetic patterns of novel lipopeptides. RESULTS: Module deletion inactivated the entire enzyme complex, whereas individual domain (A/T domain) deletion within module 7 truncated the assembly line, resulting in truncated linear hexapeptides (C(16~17)β-OHFA-Glu-Orn-Tyr-Thr-Glu-Ala/Val). Interestingly, within the module 6 catalytic unit, the effect of thiolation domain deletion differed from that of adenylation deletion. Absence of the T(6)-domain resulted in a nonproductive strain, whereas deletion of the A(6)-domain resulted in multiple assembly lines via module-skipping mechanism, generating three novel types of plipastatin derivatives, pentapeptides (C(16~17)β-OHFA-Glu-Orn-Tyr-Thr-Glu), hexapeptides (C(16~17)β-OHFA-Glu-Orn-Tyr-Thr-Glu-Ile), and octapeptides (C(16~17)β-OHFA-Glu-Orn-Tyr-Thr-Glu-Gln-Tyr-Ile). CONCLUSIONS: Notably, a unique module-skipping process occurred following deletion of the A(6)-domain, which has not been previously reported for engineered NRPS systems. This finding provides new insight into the lipopeptides engineering. It is of significant importance for combinatorial approaches and should be taken into consideration in engineering non-ribosomal peptide biosynthetic pathways for generating novel lipopeptides. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-018-0929-4) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5984369 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-59843692018-06-07 Module and individual domain deletions of NRPS to produce plipastatin derivatives in Bacillus subtilis Gao, Ling Guo, Jianping Fan, Yun Ma, Zhi Lu, Zhaoxin Zhang, Chong Zhao, Haizhen Bie, Xiaomei Microb Cell Fact Research BACKGROUND: Plipastatin, an antifungal lipopeptide, is synthesized by a non-ribosomal peptide synthetase (NRPS) in Bacillus subtilis. However, little information is available on the combinatorial biosynthesis strategies applied in plipastatin biosynthetic pathway. In this study, we applied module or individual domain deletion strategies to engineer the plipastatin biosynthetic pathway, and investigated the effect of deletions on the plipastatin assembly line, as well as revealed the synthetic patterns of novel lipopeptides. RESULTS: Module deletion inactivated the entire enzyme complex, whereas individual domain (A/T domain) deletion within module 7 truncated the assembly line, resulting in truncated linear hexapeptides (C(16~17)β-OHFA-Glu-Orn-Tyr-Thr-Glu-Ala/Val). Interestingly, within the module 6 catalytic unit, the effect of thiolation domain deletion differed from that of adenylation deletion. Absence of the T(6)-domain resulted in a nonproductive strain, whereas deletion of the A(6)-domain resulted in multiple assembly lines via module-skipping mechanism, generating three novel types of plipastatin derivatives, pentapeptides (C(16~17)β-OHFA-Glu-Orn-Tyr-Thr-Glu), hexapeptides (C(16~17)β-OHFA-Glu-Orn-Tyr-Thr-Glu-Ile), and octapeptides (C(16~17)β-OHFA-Glu-Orn-Tyr-Thr-Glu-Gln-Tyr-Ile). CONCLUSIONS: Notably, a unique module-skipping process occurred following deletion of the A(6)-domain, which has not been previously reported for engineered NRPS systems. This finding provides new insight into the lipopeptides engineering. It is of significant importance for combinatorial approaches and should be taken into consideration in engineering non-ribosomal peptide biosynthetic pathways for generating novel lipopeptides. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-018-0929-4) contains supplementary material, which is available to authorized users. BioMed Central 2018-05-31 /pmc/articles/PMC5984369/ /pubmed/29855381 http://dx.doi.org/10.1186/s12934-018-0929-4 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Gao, Ling Guo, Jianping Fan, Yun Ma, Zhi Lu, Zhaoxin Zhang, Chong Zhao, Haizhen Bie, Xiaomei Module and individual domain deletions of NRPS to produce plipastatin derivatives in Bacillus subtilis |
title | Module and individual domain deletions of NRPS to produce plipastatin derivatives in Bacillus subtilis |
title_full | Module and individual domain deletions of NRPS to produce plipastatin derivatives in Bacillus subtilis |
title_fullStr | Module and individual domain deletions of NRPS to produce plipastatin derivatives in Bacillus subtilis |
title_full_unstemmed | Module and individual domain deletions of NRPS to produce plipastatin derivatives in Bacillus subtilis |
title_short | Module and individual domain deletions of NRPS to produce plipastatin derivatives in Bacillus subtilis |
title_sort | module and individual domain deletions of nrps to produce plipastatin derivatives in bacillus subtilis |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5984369/ https://www.ncbi.nlm.nih.gov/pubmed/29855381 http://dx.doi.org/10.1186/s12934-018-0929-4 |
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