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Structure elucidation of the redox cofactor mycofactocin reveals oligo-glycosylation by MftF
Mycofactocin (MFT) is a redox cofactor belonging to the family of ribosomally synthesized and post-translationally modified peptides (RiPPs) and is involved in alcohol metabolism of mycobacteria including Mycobacterium tuberculosis. A preliminary biosynthetic model had been established by bioinforma...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Royal Society of Chemistry
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7491314/ https://www.ncbi.nlm.nih.gov/pubmed/33014324 http://dx.doi.org/10.1039/d0sc01172j |
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author | Peña-Ortiz, Luis Graça, Ana Patrícia Guo, Huijuan Braga, Daniel Köllner, Tobias G. Regestein, Lars Beemelmanns, Christine Lackner, Gerald |
author_facet | Peña-Ortiz, Luis Graça, Ana Patrícia Guo, Huijuan Braga, Daniel Köllner, Tobias G. Regestein, Lars Beemelmanns, Christine Lackner, Gerald |
author_sort | Peña-Ortiz, Luis |
collection | PubMed |
description | Mycofactocin (MFT) is a redox cofactor belonging to the family of ribosomally synthesized and post-translationally modified peptides (RiPPs) and is involved in alcohol metabolism of mycobacteria including Mycobacterium tuberculosis. A preliminary biosynthetic model had been established by bioinformatics and in vitro studies, while the structure of natural MFT and key biosynthetic steps remained elusive. Here, we report the discovery of glycosylated MFT by (13)C-labeling metabolomics and establish a model of its biosynthesis in Mycolicibacterium smegmatis. Extensive structure elucidation including NMR revealed that MFT is decorated with up to nine β-1,4-linked glucose residues including 2-O-methylglucose. Dissection of biosynthetic genes demonstrated that the oligoglycosylation is catalyzed by the glycosyltransferase MftF. Furthermore, we confirm the redox cofactor function of glycosylated MFTs by activity-based metabolic profiling using the carveol dehydrogenase LimC and show that the MFT pool expands during cultivation on ethanol. Our results will guide future studies into the biochemical functions and physiological roles of MFT in bacteria. |
format | Online Article Text |
id | pubmed-7491314 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Royal Society of Chemistry |
record_format | MEDLINE/PubMed |
spelling | pubmed-74913142020-10-02 Structure elucidation of the redox cofactor mycofactocin reveals oligo-glycosylation by MftF Peña-Ortiz, Luis Graça, Ana Patrícia Guo, Huijuan Braga, Daniel Köllner, Tobias G. Regestein, Lars Beemelmanns, Christine Lackner, Gerald Chem Sci Chemistry Mycofactocin (MFT) is a redox cofactor belonging to the family of ribosomally synthesized and post-translationally modified peptides (RiPPs) and is involved in alcohol metabolism of mycobacteria including Mycobacterium tuberculosis. A preliminary biosynthetic model had been established by bioinformatics and in vitro studies, while the structure of natural MFT and key biosynthetic steps remained elusive. Here, we report the discovery of glycosylated MFT by (13)C-labeling metabolomics and establish a model of its biosynthesis in Mycolicibacterium smegmatis. Extensive structure elucidation including NMR revealed that MFT is decorated with up to nine β-1,4-linked glucose residues including 2-O-methylglucose. Dissection of biosynthetic genes demonstrated that the oligoglycosylation is catalyzed by the glycosyltransferase MftF. Furthermore, we confirm the redox cofactor function of glycosylated MFTs by activity-based metabolic profiling using the carveol dehydrogenase LimC and show that the MFT pool expands during cultivation on ethanol. Our results will guide future studies into the biochemical functions and physiological roles of MFT in bacteria. Royal Society of Chemistry 2020-04-23 /pmc/articles/PMC7491314/ /pubmed/33014324 http://dx.doi.org/10.1039/d0sc01172j Text en This journal is © The Royal Society of Chemistry 2020 http://creativecommons.org/licenses/by/3.0/ This article is freely available. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence (CC BY 3.0) |
spellingShingle | Chemistry Peña-Ortiz, Luis Graça, Ana Patrícia Guo, Huijuan Braga, Daniel Köllner, Tobias G. Regestein, Lars Beemelmanns, Christine Lackner, Gerald Structure elucidation of the redox cofactor mycofactocin reveals oligo-glycosylation by MftF |
title | Structure elucidation of the redox cofactor mycofactocin reveals oligo-glycosylation by MftF
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title_full | Structure elucidation of the redox cofactor mycofactocin reveals oligo-glycosylation by MftF
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title_fullStr | Structure elucidation of the redox cofactor mycofactocin reveals oligo-glycosylation by MftF
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title_full_unstemmed | Structure elucidation of the redox cofactor mycofactocin reveals oligo-glycosylation by MftF
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title_short | Structure elucidation of the redox cofactor mycofactocin reveals oligo-glycosylation by MftF
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title_sort | structure elucidation of the redox cofactor mycofactocin reveals oligo-glycosylation by mftf |
topic | Chemistry |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7491314/ https://www.ncbi.nlm.nih.gov/pubmed/33014324 http://dx.doi.org/10.1039/d0sc01172j |
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