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Translocation of the thioesterase domain for the redesign of plipastatin synthetase
Non-ribosomal peptide synthetases (NRPSs) are large enzymatic complexes that catalyse the synthesis of biologically active peptides in microorganisms. Genetic engineering has recently been applied to reprogram NRPSs to produce lipopeptides with a new sequence. The carboxyl-terminal thioesterase (TE)...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5180189/ https://www.ncbi.nlm.nih.gov/pubmed/28009004 http://dx.doi.org/10.1038/srep38467 |
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author | Gao, Ling Liu, Hongxia Ma, Zhi Han, Jinzhi Lu, Zhaoxin Dai, Chen Lv, Fengxia Bie, Xiaomei |
author_facet | Gao, Ling Liu, Hongxia Ma, Zhi Han, Jinzhi Lu, Zhaoxin Dai, Chen Lv, Fengxia Bie, Xiaomei |
author_sort | Gao, Ling |
collection | PubMed |
description | Non-ribosomal peptide synthetases (NRPSs) are large enzymatic complexes that catalyse the synthesis of biologically active peptides in microorganisms. Genetic engineering has recently been applied to reprogram NRPSs to produce lipopeptides with a new sequence. The carboxyl-terminal thioesterase (TE) domains from NRPSs catalyse cleavage products by hydrolysis or complex macrocyclization. In this study, we modified plipastatin synthetase by moving the intrinsic TE region to the end of the internal thiolation (T) domains, thus generating Bacillus subtilis strains that could produce new truncated cyclic or linear peptides of the predicted sequence, which further provided an important insight into the regioselectivity of plipastatin TE. The TE was capable of recognizing and catalysing the lactone formation between (L)-Try(3) with the last few residues (L)-Pro7 and (L)-Gln8 at the C-terminus. Additionally, the unmatched linkers connecting the TE region and T domain resulted in nonproduction strains, suggesting that the native T–TE linker is necessary and sufficient for the TE domain to release the products from the hybrid enzymes. This is the first report to demonstrate truncated cyclic lipopeptides production and module skipping by simply moving the TE domain forward in an NRPS system. |
format | Online Article Text |
id | pubmed-5180189 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-51801892016-12-29 Translocation of the thioesterase domain for the redesign of plipastatin synthetase Gao, Ling Liu, Hongxia Ma, Zhi Han, Jinzhi Lu, Zhaoxin Dai, Chen Lv, Fengxia Bie, Xiaomei Sci Rep Article Non-ribosomal peptide synthetases (NRPSs) are large enzymatic complexes that catalyse the synthesis of biologically active peptides in microorganisms. Genetic engineering has recently been applied to reprogram NRPSs to produce lipopeptides with a new sequence. The carboxyl-terminal thioesterase (TE) domains from NRPSs catalyse cleavage products by hydrolysis or complex macrocyclization. In this study, we modified plipastatin synthetase by moving the intrinsic TE region to the end of the internal thiolation (T) domains, thus generating Bacillus subtilis strains that could produce new truncated cyclic or linear peptides of the predicted sequence, which further provided an important insight into the regioselectivity of plipastatin TE. The TE was capable of recognizing and catalysing the lactone formation between (L)-Try(3) with the last few residues (L)-Pro7 and (L)-Gln8 at the C-terminus. Additionally, the unmatched linkers connecting the TE region and T domain resulted in nonproduction strains, suggesting that the native T–TE linker is necessary and sufficient for the TE domain to release the products from the hybrid enzymes. This is the first report to demonstrate truncated cyclic lipopeptides production and module skipping by simply moving the TE domain forward in an NRPS system. Nature Publishing Group 2016-12-23 /pmc/articles/PMC5180189/ /pubmed/28009004 http://dx.doi.org/10.1038/srep38467 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Gao, Ling Liu, Hongxia Ma, Zhi Han, Jinzhi Lu, Zhaoxin Dai, Chen Lv, Fengxia Bie, Xiaomei Translocation of the thioesterase domain for the redesign of plipastatin synthetase |
title | Translocation of the thioesterase domain for the redesign of plipastatin synthetase |
title_full | Translocation of the thioesterase domain for the redesign of plipastatin synthetase |
title_fullStr | Translocation of the thioesterase domain for the redesign of plipastatin synthetase |
title_full_unstemmed | Translocation of the thioesterase domain for the redesign of plipastatin synthetase |
title_short | Translocation of the thioesterase domain for the redesign of plipastatin synthetase |
title_sort | translocation of the thioesterase domain for the redesign of plipastatin synthetase |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5180189/ https://www.ncbi.nlm.nih.gov/pubmed/28009004 http://dx.doi.org/10.1038/srep38467 |
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