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An Unexpected Split‐Merge Pathway in the Assembly of the Symmetric Nonribosomal Peptide Antibiotic Closthioamide
Closthioamide (CTA) is a symmetric nonribosomal peptide (NRP) comprised of two diaminopropane‐linked polythioamidated monomers. CTA is biosynthesized by Ruminiclostridium cellulolyticum via an atypical NRP synthetase (NRPS)‐independent biosynthetic pathway. Although the logic for monomer assembly wa...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7898593/ https://www.ncbi.nlm.nih.gov/pubmed/33119936 http://dx.doi.org/10.1002/anie.202011741 |
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author | Dunbar, Kyle L. Dell, Maria Molloy, Evelyn M. Büttner, Hannah Kumpfmüller, Jana Hertweck, Christian |
author_facet | Dunbar, Kyle L. Dell, Maria Molloy, Evelyn M. Büttner, Hannah Kumpfmüller, Jana Hertweck, Christian |
author_sort | Dunbar, Kyle L. |
collection | PubMed |
description | Closthioamide (CTA) is a symmetric nonribosomal peptide (NRP) comprised of two diaminopropane‐linked polythioamidated monomers. CTA is biosynthesized by Ruminiclostridium cellulolyticum via an atypical NRP synthetase (NRPS)‐independent biosynthetic pathway. Although the logic for monomer assembly was recently elucidated, the strategy for the biosynthesis and incorporation of the diamine linker remained a mystery. By means of genome editing, synthesis, and in vitro biochemical assays, we demonstrate that the final steps in CTA maturation proceed through a surprising split‐merge pathway involving the dual use of a thiotemplated intermediate. This pathway includes the first examples of an aldo‐keto reductase catalyzing the reductive release of a thiotemplated product, and of a transthioamidating transglutaminase. In addition to clarifying the remaining steps in CTA assembly, our data shed light on largely unexplored pathways for NRPS‐independent peptide biosynthesis. |
format | Online Article Text |
id | pubmed-7898593 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-78985932021-03-03 An Unexpected Split‐Merge Pathway in the Assembly of the Symmetric Nonribosomal Peptide Antibiotic Closthioamide Dunbar, Kyle L. Dell, Maria Molloy, Evelyn M. Büttner, Hannah Kumpfmüller, Jana Hertweck, Christian Angew Chem Int Ed Engl Research Articles Closthioamide (CTA) is a symmetric nonribosomal peptide (NRP) comprised of two diaminopropane‐linked polythioamidated monomers. CTA is biosynthesized by Ruminiclostridium cellulolyticum via an atypical NRP synthetase (NRPS)‐independent biosynthetic pathway. Although the logic for monomer assembly was recently elucidated, the strategy for the biosynthesis and incorporation of the diamine linker remained a mystery. By means of genome editing, synthesis, and in vitro biochemical assays, we demonstrate that the final steps in CTA maturation proceed through a surprising split‐merge pathway involving the dual use of a thiotemplated intermediate. This pathway includes the first examples of an aldo‐keto reductase catalyzing the reductive release of a thiotemplated product, and of a transthioamidating transglutaminase. In addition to clarifying the remaining steps in CTA assembly, our data shed light on largely unexplored pathways for NRPS‐independent peptide biosynthesis. John Wiley and Sons Inc. 2020-12-23 2021-02-19 /pmc/articles/PMC7898593/ /pubmed/33119936 http://dx.doi.org/10.1002/anie.202011741 Text en © 2020 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. |
spellingShingle | Research Articles Dunbar, Kyle L. Dell, Maria Molloy, Evelyn M. Büttner, Hannah Kumpfmüller, Jana Hertweck, Christian An Unexpected Split‐Merge Pathway in the Assembly of the Symmetric Nonribosomal Peptide Antibiotic Closthioamide |
title | An Unexpected Split‐Merge Pathway in the Assembly of the Symmetric Nonribosomal Peptide Antibiotic Closthioamide |
title_full | An Unexpected Split‐Merge Pathway in the Assembly of the Symmetric Nonribosomal Peptide Antibiotic Closthioamide |
title_fullStr | An Unexpected Split‐Merge Pathway in the Assembly of the Symmetric Nonribosomal Peptide Antibiotic Closthioamide |
title_full_unstemmed | An Unexpected Split‐Merge Pathway in the Assembly of the Symmetric Nonribosomal Peptide Antibiotic Closthioamide |
title_short | An Unexpected Split‐Merge Pathway in the Assembly of the Symmetric Nonribosomal Peptide Antibiotic Closthioamide |
title_sort | unexpected split‐merge pathway in the assembly of the symmetric nonribosomal peptide antibiotic closthioamide |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7898593/ https://www.ncbi.nlm.nih.gov/pubmed/33119936 http://dx.doi.org/10.1002/anie.202011741 |
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