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Heterologous Expression of a Cryptic Gene Cluster from Streptomyces leeuwenhoekii C34(T) Yields a Novel Lasso Peptide, Leepeptin

Analysis of the genome sequence of Streptomyces leeuwenhoekii C34(T) identified biosynthetic gene clusters (BGCs) for three different lasso peptides (Lp1, Lp2, and Lp3) which were not known to be made by the strain. Lasso peptides represent relatively new members of the RiPP (ribosomally synthesized...

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Autores principales: Gomez-Escribano, Juan Pablo, Castro, Jean Franco, Razmilic, Valeria, Jarmusch, Scott A., Saalbach, Gerhard, Ebel, Rainer, Jaspars, Marcel, Andrews, Barbara, Asenjo, Juan A., Bibb, Mervyn J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6856326/
https://www.ncbi.nlm.nih.gov/pubmed/31562169
http://dx.doi.org/10.1128/AEM.01752-19
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author Gomez-Escribano, Juan Pablo
Castro, Jean Franco
Razmilic, Valeria
Jarmusch, Scott A.
Saalbach, Gerhard
Ebel, Rainer
Jaspars, Marcel
Andrews, Barbara
Asenjo, Juan A.
Bibb, Mervyn J.
author_facet Gomez-Escribano, Juan Pablo
Castro, Jean Franco
Razmilic, Valeria
Jarmusch, Scott A.
Saalbach, Gerhard
Ebel, Rainer
Jaspars, Marcel
Andrews, Barbara
Asenjo, Juan A.
Bibb, Mervyn J.
author_sort Gomez-Escribano, Juan Pablo
collection PubMed
description Analysis of the genome sequence of Streptomyces leeuwenhoekii C34(T) identified biosynthetic gene clusters (BGCs) for three different lasso peptides (Lp1, Lp2, and Lp3) which were not known to be made by the strain. Lasso peptides represent relatively new members of the RiPP (ribosomally synthesized and posttranslationally modified peptides) family of natural products and have not been extensively studied. Lp3, whose production could be detected in culture supernatants from S. leeuwenhoekii C34(T) and after heterologous expression of its BGC in Streptomyces coelicolor, is identical to the previously characterized chaxapeptin. Lp1, whose production could not be detected or achieved heterologously, appears to be identical to a recently identified member of the citrulassin family of lasso peptides. Since production of Lp2 by S. leeuwenhoekii C34(T) was not observed, its BGC was also expressed in S. coelicolor. The lasso peptide was isolated and its structure confirmed by mass spectrometry and nuclear magnetic resonance analyses, revealing a novel structure that appears to represent a new family of lasso peptides. IMPORTANCE Recent developments in genome sequencing combined with bioinformatic analysis have revealed that actinomycetes contain a plethora of unexpected BGCs and thus have the potential to produce many more natural products than previously thought. This reflects the inability to detect the production of these compounds under laboratory conditions, perhaps through the use of inappropriate growth media or the absence of the environmental cues required to elicit expression of the corresponding BGCs. One approach to overcoming this problem is to circumvent the regulatory mechanisms that control expression of the BGC in its natural host by deploying heterologous expression. The generally compact nature of lasso peptide BGCs makes them particularly amenable to this approach, and, in the example given here, analysis revealed a new member of the lasso peptide family of RiPPs. This approach should be readily applicable to other cryptic lasso peptide gene clusters and would also facilitate the design and production of nonnatural variants by changing the sequence encoding the core peptide, as has been achieved with other classes of RiPPs.
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spelling pubmed-68563262019-11-25 Heterologous Expression of a Cryptic Gene Cluster from Streptomyces leeuwenhoekii C34(T) Yields a Novel Lasso Peptide, Leepeptin Gomez-Escribano, Juan Pablo Castro, Jean Franco Razmilic, Valeria Jarmusch, Scott A. Saalbach, Gerhard Ebel, Rainer Jaspars, Marcel Andrews, Barbara Asenjo, Juan A. Bibb, Mervyn J. Appl Environ Microbiol Genetics and Molecular Biology Analysis of the genome sequence of Streptomyces leeuwenhoekii C34(T) identified biosynthetic gene clusters (BGCs) for three different lasso peptides (Lp1, Lp2, and Lp3) which were not known to be made by the strain. Lasso peptides represent relatively new members of the RiPP (ribosomally synthesized and posttranslationally modified peptides) family of natural products and have not been extensively studied. Lp3, whose production could be detected in culture supernatants from S. leeuwenhoekii C34(T) and after heterologous expression of its BGC in Streptomyces coelicolor, is identical to the previously characterized chaxapeptin. Lp1, whose production could not be detected or achieved heterologously, appears to be identical to a recently identified member of the citrulassin family of lasso peptides. Since production of Lp2 by S. leeuwenhoekii C34(T) was not observed, its BGC was also expressed in S. coelicolor. The lasso peptide was isolated and its structure confirmed by mass spectrometry and nuclear magnetic resonance analyses, revealing a novel structure that appears to represent a new family of lasso peptides. IMPORTANCE Recent developments in genome sequencing combined with bioinformatic analysis have revealed that actinomycetes contain a plethora of unexpected BGCs and thus have the potential to produce many more natural products than previously thought. This reflects the inability to detect the production of these compounds under laboratory conditions, perhaps through the use of inappropriate growth media or the absence of the environmental cues required to elicit expression of the corresponding BGCs. One approach to overcoming this problem is to circumvent the regulatory mechanisms that control expression of the BGC in its natural host by deploying heterologous expression. The generally compact nature of lasso peptide BGCs makes them particularly amenable to this approach, and, in the example given here, analysis revealed a new member of the lasso peptide family of RiPPs. This approach should be readily applicable to other cryptic lasso peptide gene clusters and would also facilitate the design and production of nonnatural variants by changing the sequence encoding the core peptide, as has been achieved with other classes of RiPPs. American Society for Microbiology 2019-11-14 /pmc/articles/PMC6856326/ /pubmed/31562169 http://dx.doi.org/10.1128/AEM.01752-19 Text en Copyright © 2019 Gomez-Escribano et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Genetics and Molecular Biology
Gomez-Escribano, Juan Pablo
Castro, Jean Franco
Razmilic, Valeria
Jarmusch, Scott A.
Saalbach, Gerhard
Ebel, Rainer
Jaspars, Marcel
Andrews, Barbara
Asenjo, Juan A.
Bibb, Mervyn J.
Heterologous Expression of a Cryptic Gene Cluster from Streptomyces leeuwenhoekii C34(T) Yields a Novel Lasso Peptide, Leepeptin
title Heterologous Expression of a Cryptic Gene Cluster from Streptomyces leeuwenhoekii C34(T) Yields a Novel Lasso Peptide, Leepeptin
title_full Heterologous Expression of a Cryptic Gene Cluster from Streptomyces leeuwenhoekii C34(T) Yields a Novel Lasso Peptide, Leepeptin
title_fullStr Heterologous Expression of a Cryptic Gene Cluster from Streptomyces leeuwenhoekii C34(T) Yields a Novel Lasso Peptide, Leepeptin
title_full_unstemmed Heterologous Expression of a Cryptic Gene Cluster from Streptomyces leeuwenhoekii C34(T) Yields a Novel Lasso Peptide, Leepeptin
title_short Heterologous Expression of a Cryptic Gene Cluster from Streptomyces leeuwenhoekii C34(T) Yields a Novel Lasso Peptide, Leepeptin
title_sort heterologous expression of a cryptic gene cluster from streptomyces leeuwenhoekii c34(t) yields a novel lasso peptide, leepeptin
topic Genetics and Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6856326/
https://www.ncbi.nlm.nih.gov/pubmed/31562169
http://dx.doi.org/10.1128/AEM.01752-19
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