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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...

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Autores principales: Gao, Ling, Guo, Jianping, Fan, Yun, Ma, Zhi, Lu, Zhaoxin, Zhang, Chong, Zhao, Haizhen, Bie, Xiaomei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
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.
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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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