Molecular basis for the biosynthesis of the siderophore coprogen in the cheese-ripening fungus Penicillium roqueforti

Abstract Background Iron is an essential nutrient for microorganisms, including fungi, which have evolved strategies to acquire it. The most common strategy is the secretion of siderophores, low-molecular-weight compounds with a high affinity for ferric ions, which are involved in cellular iron upta...

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Main Authors: Kathia González, Mariana Montanares, Matías Gallardo, Carlos Gil-Durán, Abel M. Forero, Jaime Rodríguez, Carlos Jiménez, Inmaculada Vaca, Renato Chávez
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Biological Research
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Online Access:https://doi.org/10.1186/s40659-025-00633-2
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author Kathia González
Mariana Montanares
Matías Gallardo
Carlos Gil-Durán
Abel M. Forero
Jaime Rodríguez
Carlos Jiménez
Inmaculada Vaca
Renato Chávez
author_facet Kathia González
Mariana Montanares
Matías Gallardo
Carlos Gil-Durán
Abel M. Forero
Jaime Rodríguez
Carlos Jiménez
Inmaculada Vaca
Renato Chávez
author_sort Kathia González
collection DOAJ
description Abstract Background Iron is an essential nutrient for microorganisms, including fungi, which have evolved strategies to acquire it. The most common strategy is the secretion of siderophores, low-molecular-weight compounds with a high affinity for ferric ions, which are involved in cellular iron uptake. Penicillium roqueforti, the fungus responsible for the ripening of blue-veined cheeses, produces coprogen, a hydroxamate-type siderophore. However, to date, the molecular basis for its biosynthesis remains elusive. Results In this study, we identified and characterized a biosynthetic gene cluster (BGC) responsible for coprogen biosynthesis in P. roqueforti, named the cop BGC. This BGC contains seven genes, three of which (copA, copB and copE) encode enzymes directly involved in coprogen biosynthesis from precursors molecules. Using CRISPR-Cas9, we targeted these three genes and analyzed the resulting mutants by Liquid Chromatography/High-Resolution Mass Spectrometry (LC/HRMS). Our results confirmed that all three genes are necessary for coprogen biosynthesis. Phenotypically, the mutants displayed growth differences under iron-deficient conditions, which correlated with their ability to either synthesize or fail to synthesize coprogen B and dimerumic acid, intermediates in the coprogen pathway with siderophore activity. Conclusions The results obtained in this work provide important insights into the molecular basis of coprogen biosynthesis in P. roqueforti, enhancing the understanding of how siderophores enable this fungus to thrive in iron-deficient environments.
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spelling doaj-art-da2ddeca78fc44d2af622005fed3b2372025-08-20T03:04:10ZengBMCBiological Research0717-62872025-07-0158111410.1186/s40659-025-00633-2Molecular basis for the biosynthesis of the siderophore coprogen in the cheese-ripening fungus Penicillium roquefortiKathia González0Mariana Montanares1Matías Gallardo2Carlos Gil-Durán3Abel M. Forero4Jaime Rodríguez5Carlos Jiménez6Inmaculada Vaca7Renato Chávez8Departamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile (USACH)Departamento de Química, Facultad de Ciencias, Universidad de ChileDepartamento de Química, Facultad de Ciencias, Universidad de ChileDepartamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile (USACH)CICA-Centro Interdisciplinar de Química e Bioloxía, Departamento de Química, Facultade de Ciencias, Universidade da CoruñaCICA-Centro Interdisciplinar de Química e Bioloxía, Departamento de Química, Facultade de Ciencias, Universidade da CoruñaCICA-Centro Interdisciplinar de Química e Bioloxía, Departamento de Química, Facultade de Ciencias, Universidade da CoruñaDepartamento de Química, Facultad de Ciencias, Universidad de ChileDepartamento de Biología, Facultad de Química y Biología, Universidad de Santiago de Chile (USACH)Abstract Background Iron is an essential nutrient for microorganisms, including fungi, which have evolved strategies to acquire it. The most common strategy is the secretion of siderophores, low-molecular-weight compounds with a high affinity for ferric ions, which are involved in cellular iron uptake. Penicillium roqueforti, the fungus responsible for the ripening of blue-veined cheeses, produces coprogen, a hydroxamate-type siderophore. However, to date, the molecular basis for its biosynthesis remains elusive. Results In this study, we identified and characterized a biosynthetic gene cluster (BGC) responsible for coprogen biosynthesis in P. roqueforti, named the cop BGC. This BGC contains seven genes, three of which (copA, copB and copE) encode enzymes directly involved in coprogen biosynthesis from precursors molecules. Using CRISPR-Cas9, we targeted these three genes and analyzed the resulting mutants by Liquid Chromatography/High-Resolution Mass Spectrometry (LC/HRMS). Our results confirmed that all three genes are necessary for coprogen biosynthesis. Phenotypically, the mutants displayed growth differences under iron-deficient conditions, which correlated with their ability to either synthesize or fail to synthesize coprogen B and dimerumic acid, intermediates in the coprogen pathway with siderophore activity. Conclusions The results obtained in this work provide important insights into the molecular basis of coprogen biosynthesis in P. roqueforti, enhancing the understanding of how siderophores enable this fungus to thrive in iron-deficient environments.https://doi.org/10.1186/s40659-025-00633-2Penicillium roquefortiSiderophoreCoprogenBiosynthetic gene clusterCRISPR-Cas9Mass spectrometry
spellingShingle Kathia González
Mariana Montanares
Matías Gallardo
Carlos Gil-Durán
Abel M. Forero
Jaime Rodríguez
Carlos Jiménez
Inmaculada Vaca
Renato Chávez
Molecular basis for the biosynthesis of the siderophore coprogen in the cheese-ripening fungus Penicillium roqueforti
Biological Research
Penicillium roqueforti
Siderophore
Coprogen
Biosynthetic gene cluster
CRISPR-Cas9
Mass spectrometry
title Molecular basis for the biosynthesis of the siderophore coprogen in the cheese-ripening fungus Penicillium roqueforti
title_full Molecular basis for the biosynthesis of the siderophore coprogen in the cheese-ripening fungus Penicillium roqueforti
title_fullStr Molecular basis for the biosynthesis of the siderophore coprogen in the cheese-ripening fungus Penicillium roqueforti
title_full_unstemmed Molecular basis for the biosynthesis of the siderophore coprogen in the cheese-ripening fungus Penicillium roqueforti
title_short Molecular basis for the biosynthesis of the siderophore coprogen in the cheese-ripening fungus Penicillium roqueforti
title_sort molecular basis for the biosynthesis of the siderophore coprogen in the cheese ripening fungus penicillium roqueforti
topic Penicillium roqueforti
Siderophore
Coprogen
Biosynthetic gene cluster
CRISPR-Cas9
Mass spectrometry
url https://doi.org/10.1186/s40659-025-00633-2
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