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Sylvie Lautru - One of the best experts on this subject based on the ideXlab platform.

  • Natural Combinatorial Biosynthesis Involving Two Clusters for the Synthesis of Three Pyrrolamides in Streptomyces netropsis.
    ACS Chemical Biology, 2015
    Co-Authors: Audrey Vingadassalon, Florence Lorieux, Maud Juguet, Géraldine Le Goff, Claude Gerbaud, Jean-luc Pernodet, Sylvie Lautru
    Abstract:

    The pyrrolamides constitute a small family of secondary metabolites that are known for their ability to bind noncovalently to the DNA minor groove with some sequence specificity. To date, only a single pyrrolamide biosynthetic gene cluster has been reported, directing the synthesis of Congocidine (netropsin) in Streptomyces ambofaciens. In this study, we improve our understanding of pyrrolamide biosynthesis through the identification and characterization of the gene cluster responsible for the production of distamycin in Streptomyces netropsis DSM40846. We discover that the strain produces two other pyrrolamides, the well-characterized Congocidine and a Congocidine/distamycin hybrid that we named disgocidine. S. netropsis DSM40846 genome analysis led to the identification of two distinct pyrrolamide-like biosynthetic gene clusters. We show here that these two clusters are reciprocally dependent for the production of the three pyrrolamide molecules. Furthermore, based on detailed functional analysis of these clusters, we propose a biosynthetic route to Congocidine and distamycin and an updated model for pyrrolamide assembly. The synthesis of disgocidine, the distamycin/Congocidine hybrid, appears to constitute the first example of "natural combinatorial biosynthesis" between two related biosynthetic pathways. Finally, we analyze the genomic context of the two biosynthetic gene clusters and suggest that the presently interdependent clusters result from the coevolution of two ancestral independent pyrrolamide gene clusters.

  • Natural Combinatorial Biosynthesis Involving Two Clusters for the Synthesis of Three Pyrrolamides in Streptomyces netropsis
    2015
    Co-Authors: Audrey Vingadassalon, Florence Lorieux, Maud Juguet, Claude Gerbaud, Jean-luc Pernodet, Géraldine Le Goff, Sylvie Lautru
    Abstract:

    The pyrrolamides constitute a small family of secondary metabolites that are known for their ability to bind noncovalently to the DNA minor groove with some sequence specificity. To date, only a single pyrrolamide biosynthetic gene cluster has been reported, directing the synthesis of Congocidine (netropsin) in Streptomyces ambofaciens. In this study, we improve our understanding of pyrrolamide biosynthesis through the identification and characterization of the gene cluster responsible for the production of distamycin in Streptomyces netropsis DSM40846. We discover that the strain produces two other pyrrolamides, the well-characterized Congocidine and a Congocidine/distamycin hybrid that we named disgocidine. S. netropsis DSM40846 genome analysis led to the identification of two distinct pyrrolamide-like biosynthetic gene clusters. We show here that these two clusters are reciprocally dependent for the production of the three pyrrolamide molecules. Furthermore, based on detailed functional analysis of these clusters, we propose a biosynthetic route to Congocidine and distamycin and an updated model for pyrrolamide assembly. The synthesis of disgocidine, the distamycin/Congocidine hybrid, appears to constitute the first example of “natural combinatorial biosynthesis” between two related biosynthetic pathways. Finally, we analyze the genomic context of the two biosynthetic gene clusters and suggest that the presently interdependent clusters result from the coevolution of two ancestral independent pyrrolamide gene clusters

  • Genome mining of Streptomyces ambofaciens
    Journal of Industrial Microbiology and Biotechnology, 2014
    Co-Authors: Bertrand Aigle, Pierre Leblond, Sylvie Lautru, Dieter Spiteller, Jeroen S. Dickschat, Gregory L. Challis, Jean-luc Pernodet
    Abstract:

    Since the discovery of the streptomycin produced by Streptomyces griseus in the middle of the last century, members of this bacterial genus have been largely exploited for the production of secondary metabolites with wide uses in medicine and in agriculture. They have even been recognized as one of the most prolific producers of natural products among microorganisms. With the onset of the genomic era, it became evident that these microorganisms still represent a major source for the discovery of novel secondary metabolites. This was highlighted with the complete genome sequencing of Streptomyces coelicolor A3(2) which revealed an unexpected potential of this organism to synthesize natural products undetected until then by classical screening methods. Since then, analysis of sequenced genomes from numerous Streptomyces species has shown that a single species can carry more than 30 secondary metabolite gene clusters, reinforcing the idea that the biosynthetic potential of this bacterial genus is far from being fully exploited. This review highlights our knowledge on the potential of Streptomyces ambofaciens ATCC 23877 to synthesize natural products. This industrial strain was known for decades to only produce the drug spiramycin and another antibacterial compound, Congocidine. Mining of its genome allowed the identification of 23 clusters potentially involved in the production of other secondary metabolites. Studies of some of these clusters resulted in the characterization of novel compounds and of previously known compounds but never characterized in this Streptomyces species. In addition, genome mining revealed that secondary metabolite gene clusters of phylogenetically closely related Streptomyces are mainly species-specific.

  • Genome-guided Exploration of Streptomyces ambofaciens Secondary Metabolism
    2011
    Co-Authors: Bertrand Aigle, Sylvie Lautru, Sarka Nezbedova, Robert Bunet, Christophe Corre, Amélie Garenaux, S. Huang, Luisa Laureti, Maria Vaz Mendez, H.c. Nguyen
    Abstract:

    Members of the Streptomyces genus are among the most prolific microorganisms producing secondary metabolites with wide uses in medicine and in agriculture. Sequencing of the genome of the model Streptomyces, Streptomyces coelicolor, has highlighted an unexpected feature, i.e. that the potential of these organisms to synthesise secondary metabolites has been largely underestimated. They indeed possess many more gene clusters encoding natural product-like biosynthetic pathways than there are known natural products. Similar observations have since been made for other bacterial or fungal genomes. Thus, it became clear that microbial secondary metabolism had been seriously underestimated and that genome-based approaches were very promising for the search of new bioactive compounds. Here, we present an overview of the secondary metabolite biosynthetic potential of Streptomyces ambofaciens, a species known for decades as producer of the macrolide spiramycin and the pyrrolamide Congocidine. Interestingly, genome analysis has revealed that despite of the close phylogenetic relatedness between S. coelicolor and S. ambofaciens, most of its secondary metabolite gene clusters are species-specific.

  • an iterative nonribosomal peptide synthetase assembles the pyrrole amide antibiotic Congocidine in streptomyces ambofaciens
    Chemistry & Biology, 2009
    Co-Authors: Maud Juguet, Pierre Leblond, Sylvie Lautru, Francoisxavier Francou, Sarka Nezbedova, Muriel Gondry, Jean-luc Pernodet
    Abstract:

    Congocidine (netropsin) is a pyrrole-amide (oligopyrrole, oligopeptide) antibiotic produced by Streptomyces ambofaciens. We have identified, in the right terminal region of the S. ambofaciens chromosome, the gene cluster that directs Congocidine biosynthesis. Heterologous expression of the cluster and in-frame deletions of 8 of the 22 genes confirm the involvement of this cluster in Congocidine biosynthesis. Nine genes can be assigned specific functions in regulation, resistance, or Congocidine assembly. In contrast, the biosynthetic origin of the precursors cannot be easily inferred from in silico analyses. Congocidine is assembled by a nonribosomal peptide synthetase (NRPS) constituted of a free-standing module and several single-domain proteins encoded by four genes. The iterative use of its unique adenylation domain, the utilization of guanidinoacetyl-CoA as a substrate by a condensation domain, and the control of 4-aminopyrrole-2-carboxylate polymerization constitute the most original features of this NRPS.

Jean-luc Pernodet - One of the best experts on this subject based on the ideXlab platform.

  • Natural Combinatorial Biosynthesis Involving Two Clusters for the Synthesis of Three Pyrrolamides in Streptomyces netropsis.
    ACS Chemical Biology, 2015
    Co-Authors: Audrey Vingadassalon, Florence Lorieux, Maud Juguet, Géraldine Le Goff, Claude Gerbaud, Jean-luc Pernodet, Sylvie Lautru
    Abstract:

    The pyrrolamides constitute a small family of secondary metabolites that are known for their ability to bind noncovalently to the DNA minor groove with some sequence specificity. To date, only a single pyrrolamide biosynthetic gene cluster has been reported, directing the synthesis of Congocidine (netropsin) in Streptomyces ambofaciens. In this study, we improve our understanding of pyrrolamide biosynthesis through the identification and characterization of the gene cluster responsible for the production of distamycin in Streptomyces netropsis DSM40846. We discover that the strain produces two other pyrrolamides, the well-characterized Congocidine and a Congocidine/distamycin hybrid that we named disgocidine. S. netropsis DSM40846 genome analysis led to the identification of two distinct pyrrolamide-like biosynthetic gene clusters. We show here that these two clusters are reciprocally dependent for the production of the three pyrrolamide molecules. Furthermore, based on detailed functional analysis of these clusters, we propose a biosynthetic route to Congocidine and distamycin and an updated model for pyrrolamide assembly. The synthesis of disgocidine, the distamycin/Congocidine hybrid, appears to constitute the first example of "natural combinatorial biosynthesis" between two related biosynthetic pathways. Finally, we analyze the genomic context of the two biosynthetic gene clusters and suggest that the presently interdependent clusters result from the coevolution of two ancestral independent pyrrolamide gene clusters.

  • Natural Combinatorial Biosynthesis Involving Two Clusters for the Synthesis of Three Pyrrolamides in Streptomyces netropsis
    2015
    Co-Authors: Audrey Vingadassalon, Florence Lorieux, Maud Juguet, Claude Gerbaud, Jean-luc Pernodet, Géraldine Le Goff, Sylvie Lautru
    Abstract:

    The pyrrolamides constitute a small family of secondary metabolites that are known for their ability to bind noncovalently to the DNA minor groove with some sequence specificity. To date, only a single pyrrolamide biosynthetic gene cluster has been reported, directing the synthesis of Congocidine (netropsin) in Streptomyces ambofaciens. In this study, we improve our understanding of pyrrolamide biosynthesis through the identification and characterization of the gene cluster responsible for the production of distamycin in Streptomyces netropsis DSM40846. We discover that the strain produces two other pyrrolamides, the well-characterized Congocidine and a Congocidine/distamycin hybrid that we named disgocidine. S. netropsis DSM40846 genome analysis led to the identification of two distinct pyrrolamide-like biosynthetic gene clusters. We show here that these two clusters are reciprocally dependent for the production of the three pyrrolamide molecules. Furthermore, based on detailed functional analysis of these clusters, we propose a biosynthetic route to Congocidine and distamycin and an updated model for pyrrolamide assembly. The synthesis of disgocidine, the distamycin/Congocidine hybrid, appears to constitute the first example of “natural combinatorial biosynthesis” between two related biosynthetic pathways. Finally, we analyze the genomic context of the two biosynthetic gene clusters and suggest that the presently interdependent clusters result from the coevolution of two ancestral independent pyrrolamide gene clusters

  • Genome mining of Streptomyces ambofaciens
    Journal of Industrial Microbiology and Biotechnology, 2014
    Co-Authors: Bertrand Aigle, Pierre Leblond, Sylvie Lautru, Dieter Spiteller, Jeroen S. Dickschat, Gregory L. Challis, Jean-luc Pernodet
    Abstract:

    Since the discovery of the streptomycin produced by Streptomyces griseus in the middle of the last century, members of this bacterial genus have been largely exploited for the production of secondary metabolites with wide uses in medicine and in agriculture. They have even been recognized as one of the most prolific producers of natural products among microorganisms. With the onset of the genomic era, it became evident that these microorganisms still represent a major source for the discovery of novel secondary metabolites. This was highlighted with the complete genome sequencing of Streptomyces coelicolor A3(2) which revealed an unexpected potential of this organism to synthesize natural products undetected until then by classical screening methods. Since then, analysis of sequenced genomes from numerous Streptomyces species has shown that a single species can carry more than 30 secondary metabolite gene clusters, reinforcing the idea that the biosynthetic potential of this bacterial genus is far from being fully exploited. This review highlights our knowledge on the potential of Streptomyces ambofaciens ATCC 23877 to synthesize natural products. This industrial strain was known for decades to only produce the drug spiramycin and another antibacterial compound, Congocidine. Mining of its genome allowed the identification of 23 clusters potentially involved in the production of other secondary metabolites. Studies of some of these clusters resulted in the characterization of novel compounds and of previously known compounds but never characterized in this Streptomyces species. In addition, genome mining revealed that secondary metabolite gene clusters of phylogenetically closely related Streptomyces are mainly species-specific.

  • an iterative nonribosomal peptide synthetase assembles the pyrrole amide antibiotic Congocidine in streptomyces ambofaciens
    Chemistry & Biology, 2009
    Co-Authors: Maud Juguet, Pierre Leblond, Sylvie Lautru, Francoisxavier Francou, Sarka Nezbedova, Muriel Gondry, Jean-luc Pernodet
    Abstract:

    Congocidine (netropsin) is a pyrrole-amide (oligopyrrole, oligopeptide) antibiotic produced by Streptomyces ambofaciens. We have identified, in the right terminal region of the S. ambofaciens chromosome, the gene cluster that directs Congocidine biosynthesis. Heterologous expression of the cluster and in-frame deletions of 8 of the 22 genes confirm the involvement of this cluster in Congocidine biosynthesis. Nine genes can be assigned specific functions in regulation, resistance, or Congocidine assembly. In contrast, the biosynthetic origin of the precursors cannot be easily inferred from in silico analyses. Congocidine is assembled by a nonribosomal peptide synthetase (NRPS) constituted of a free-standing module and several single-domain proteins encoded by four genes. The iterative use of its unique adenylation domain, the utilization of guanidinoacetyl-CoA as a substrate by a condensation domain, and the control of 4-aminopyrrole-2-carboxylate polymerization constitute the most original features of this NRPS.

Pernodet Jean-luc - One of the best experts on this subject based on the ideXlab platform.

  • Revised structure of anthelvencin A and characterization of the anthelvencin biosynthetic gene cluster
    'American Chemical Society (ACS)', 2020
    Co-Authors: Aubry Céline, Pernodet Jean-luc, Gerbaud Claude, Clerici Paolo, Micouin Laurent, Lautru Sylvie
    Abstract:

    International audienceAnthelvencins A and B are pyrrolamide metabolites produced by Streptomyces venezuelae ATCC 14583 and 14585. Isolated in 1965, they were reported to exhibit anthelmintic and moderate antibacterial activities. In this study, we revise the structure of anthelvencin A and identify a third anthelvencin metabolite, bearing two N-methylated pyrrole groups, which we named anthelvencin C. We sequenced the genome of S. venezuelae ATCC 14583 and identified a gene cluster predicted to direct the biosynthesis of anthelvencins. Functional analysis of this gene cluster confirmed its involvement in anthelvencin biosynthesis and allowed us to propose a biosynthetic pathway for anthelvencins. In addition to a non-ribosomal peptide synthetase (NRPS), the assembly of anthelvencins involves an enzyme from the ATP-grasp ligase family, Ant23. We propose that Ant23 uses a PCP-loaded 4-aminopyrrole-2-carboxylate as substrate. As observed for the biosynthesis of the other pyrrolamides Congocidine (produced by Streptomyces ambofaciens ATCC 25877) and distamycin (produced by Streptomyces netropsis DSM 40846), the NRPS assembling anthelvencins is composed of stand-alone domains only. Such NRPSs, sometimes called type II NRPSs, are less studied than the classical multimodular NRPSs. Yet, they constitute an interesting model to study protein-protein interactions in NRPSs and are good candidates for combinatorial biosynthesis approaches

  • Natural Combinatorial Biosynthesis Involving Two Clusters for the Synthesis of Three Pyrrolamides in Streptomyces netropsis.
    'American Chemical Society (ACS)', 2015
    Co-Authors: Vingadassalon Audrey, Pernodet Jean-luc, Juguet Maud, Lorieux Florence, Le Goff Géraldine, Gerbaud Claude, Lautru Sylvie
    Abstract:

    International audienceThe pyrrolamides constitute a small family of secondary metabolites that are known for their ability to bind noncovalently to the DNA minor groove with some sequence specificity. To date, only a single pyrrolamide biosynthetic gene cluster has been reported, directing the synthesis of Congocidine (netropsin) in Streptomyces ambofaciens. In this study, we improve our understanding of pyrrolamide biosynthesis through the identification and characterization of the gene cluster responsible for the production of distamycin in Streptomyces netropsis DSM40846. We discover that the strain produces two other pyrrolamides, the well-characterized Congocidine and a Congocidine/distamycin hybrid that we named disgocidine. S. netropsis DSM40846 genome analysis led to the identification of two distinct pyrrolamide-like biosynthetic gene clusters. We show here that these two clusters are reciprocally dependent for the production of the three pyrrolamide molecules. Furthermore, based on detailed functional analysis of these clusters, we propose a biosynthetic route to Congocidine and distamycin and an updated model for pyrrolamide assembly. The synthesis of disgocidine, the distamycin/Congocidine hybrid, appears to constitute the first example of "natural combinatorial biosynthesis" between two related biosynthetic pathways. Finally, we analyze the genomic context of the two biosynthetic gene clusters and suggest that the presently interdependent clusters result from the coevolution of two ancestral independent pyrrolamide gene clusters

  • Genome mining of Streptomyces ambofaciens
    'Springer Science and Business Media LLC', 2014
    Co-Authors: Aigle Bertrand, Leblond Pierre, Lautru Sylvie, Spiteller Dieter, Dickschat, Jeroen S., Challis, Gregory L., Pernodet Jean-luc
    Abstract:

    International audienceSince the discovery of the streptomycin produced by Streptomyces griseus in the middle of the last century, members of this bacterial genus have been largely exploited for the production of secondary metabolites with wide uses in medicine and in agriculture. They have even been recognized as one of the most prolific producers of natural products among microorganisms. With the onset of the genomic era, it became evident that these microorganisms still represent a major source for the discovery of novel secondary metabolites. This was highlighted with the complete genome sequencing of Streptomyces coelicolor A3(2) which revealed an unexpected potential of this organism to synthesize natural products undetected until then by classical screening methods. Since then, analysis of sequenced genomes from numerous Streptomyces species has shown that a single species can carry more than 30 secondary metabolite gene clusters, reinforcing the idea that the biosynthetic potential of this bacterial genus is far from being fully exploited. This review highlights our knowledge on the potential of Streptomyces ambofaciens ATCC 23877 to synthesize natural products. This industrial strain was known for decades to only produce the drug spiramycin and another antibacterial compound, Congocidine. Mining of its genome allowed the identification of 23 clusters potentially involved in the production of other secondary metabolites. Studies of some of these clusters resulted in the characterization of novel compounds and of previously known compounds but never characterized in this Streptomyces species. In addition, genome mining revealed that secondary metabolite gene clusters of phylogenetically closely related Streptomyces are mainly species-specific

  • Caractérisation de voies de biosynthèse d'antibiotiques de la famille des pyrrolamides
    2013
    Co-Authors: Vingadassalon Audrey, Pernodet Jean-luc
    Abstract:

    Les pyrrolamides constituent une famille de produits naturels dotés de diverses activités biologiques et synthétisés par des actinobactéries. La Congocidine et la distamycine, les molécules les plus connues de cette famille, sont capables de se lier à l'ADN de façon non covalente selon une certaine spécificité de séquence (succession de 4 paires de base A/T). Récemment, les gènes et la voie de biosynthèse de la Congocidine ont été identifiés et caractérisés chez S. ambofaciens. Ceci a révélé un mécanisme original impliquant notamment de nouvelles enzymes et de nouvelles voies pour la biosynthèse des trois précurseurs nécessaires à l assemblage de la Congocidine. Nous avons entrepris d étudier la régulation de la biosynthèse de la Congocidine chez S. ambofaciens et d isoler et de caractériser les groupes de gènes de biosynthèse de deux autres pyrrolamides, la distamycine et les pyrronamycines (produites respectivement par S distallicus et un streptomyces non caractérisé). L'objectif de cette étude est, dans un premier temps, d améliorer notre compréhension des mécanismes impliqués lors de la biosynthèse de ces molécules (comme le mécanisme d incorporation des pyrroles) et, par la suite, de manipuler les gènes identifiés pour synthétiser de nouvelles molécules pyrrolamides hybrides.Pyrrolamides constitute a family of natural products with various biological activities, synthesized by actinobacteria. Congocidine (also called netropsin) and distamycin are the best characterized pyrrolamides, largely studied due to their ability to bind into the minor groove of the DNA double helix in a sequence specific manner (succession of four A/T bases). Recently, the Congocidine biosynthetic pathway has been characterized in Streptomyces ambofaciens. We showed that an iterative Non Ribosomal Peptide Synthetase with an unusual architecture assembles Congocidine, using precursors with undocumented biosynthetic pathways. With the aim of developing a combinatorial biosynthesis approach for the development of new pyrrolamides, we undertook the study of the regulation of Congocidine biosynthesis in S. ambofaciens and the isolation of the distamycin and pyrronamycins biosynthetic gene clusters. Characterization of these clusters will result in a more detailed understanding of pyrrolamide biosynthesis (e.g. mechanism of pyrrole polymerization), and provide new tools (enzymes) and building blocks (precursors) necessary for combinatorial biosynthesis.PARIS11-SCD-Bib. électronique (914719901) / SudocSudocFranceF

  • A sweet origin for the key Congocidine precursor 4-acetamidopyrrole-2-carboxylate
    'Wiley', 2012
    Co-Authors: Lautru Sylvie, Challis, Gregory L., Song Lijiang, Demange Luc, Lombès Thomas, Galons Hervé, Pernodet Jean-luc
    Abstract:

    Feeding (Streptomyces) frenzy: Natural products belonging to the pyrrolamide family are defined by their pyrrole-2-carboxamide moiety. 4-acetamidopyrrole-2-carboxylate is identified as the key pyrrolamide Congocidine precursor (see scheme) through feeding studies using Streptomyces ambofaciens. The biosynthetic pathway of Congocidine starts with the carbohydrate N-acetylglucosamine and involves carbohydrate-processing enzymes

Bertrand Aigle - One of the best experts on this subject based on the ideXlab platform.

  • Genome mining of Streptomyces ambofaciens
    Journal of Industrial Microbiology and Biotechnology, 2014
    Co-Authors: Bertrand Aigle, Pierre Leblond, Sylvie Lautru, Dieter Spiteller, Jeroen S. Dickschat, Gregory L. Challis, Jean-luc Pernodet
    Abstract:

    Since the discovery of the streptomycin produced by Streptomyces griseus in the middle of the last century, members of this bacterial genus have been largely exploited for the production of secondary metabolites with wide uses in medicine and in agriculture. They have even been recognized as one of the most prolific producers of natural products among microorganisms. With the onset of the genomic era, it became evident that these microorganisms still represent a major source for the discovery of novel secondary metabolites. This was highlighted with the complete genome sequencing of Streptomyces coelicolor A3(2) which revealed an unexpected potential of this organism to synthesize natural products undetected until then by classical screening methods. Since then, analysis of sequenced genomes from numerous Streptomyces species has shown that a single species can carry more than 30 secondary metabolite gene clusters, reinforcing the idea that the biosynthetic potential of this bacterial genus is far from being fully exploited. This review highlights our knowledge on the potential of Streptomyces ambofaciens ATCC 23877 to synthesize natural products. This industrial strain was known for decades to only produce the drug spiramycin and another antibacterial compound, Congocidine. Mining of its genome allowed the identification of 23 clusters potentially involved in the production of other secondary metabolites. Studies of some of these clusters resulted in the characterization of novel compounds and of previously known compounds but never characterized in this Streptomyces species. In addition, genome mining revealed that secondary metabolite gene clusters of phylogenetically closely related Streptomyces are mainly species-specific.

  • Effect of the deletion of sco6673-like on the spiramycin and Congocidine production.
    2014
    Co-Authors: Robert Bunet, Pierre Leblond, Luisa Laureti, Dieter Spiteller, Jeroen S. Dickschat, L. Hotel, Ramona Riclea, Cédric Paris, Jean-michel Girardet, Bertrand Aigle
    Abstract:

    (A) Spiramycin and Congocidine production was assessed directly by HPLC from supernatant samples collected from the culture of S. ambofaciens ATCC23877 wild-type and mutant strains grown in liquid MP5. Commercial spiramycin (100 µl at 0.1 mg/ml) was used as control. Absorption was monitored at 232 nm. The inserts are an enlargement of the area between 20 to 35 min of retention time containing the peaks corresponding to the spiramycin (spiramycin is a mixture of three forms). The peak corresponding to Congocidine is highlighted with a black dot. (B) UV spectra (from 200 to 350 nm) of the peaks highlighted with an asterisk and corresponding to spiramycin.

  • A Single Sfp-Type Phosphopantetheinyl Transferase Plays a Major Role in the Biosynthesis of PKS and NRPS Derived Metabolites in Streptomyces ambofaciens ATCC23877
    PLoS ONE, 2014
    Co-Authors: Robert Bunet, Pierre Leblond, Luisa Laureti, Dieter Spiteller, Jeroen S. Dickschat, L. Hotel, Ramona Riclea, Cédric Paris, Jean-michel Girardet, Bertrand Aigle
    Abstract:

    The phosphopantetheinyl transferases (PPTases) are responsible for the activation of the carrier protein domains of the polyketide synthases (PKS), non ribosomal peptide synthases (NRPS) and fatty acid synthases (FAS). The analysis of the Streptomyces ambofaciens ATCC23877 genome has revealed the presence of four putative PPTase encoding genes. One of these genes appears to be essential and is likely involved in fatty acid biosynthesis. Two other PPTase genes, samT0172 (alpN) and samL0372, are located within a type II PKS gene cluster responsible for the kinamycin production and an hybrid NRPS-PKS cluster involved in antimycin production, respectively, and their products were shown to be specifically involved in the biosynthesis of these secondary metabolites. Surprisingly, the fourth PPTase gene, which is not located within a secondary metabolite gene cluster, appears to play a pleiotropic role. Its product is likely involved in the activation of the acyl- and peptidyl-carrier protein domains within all the other PKS and NRPS complexes encoded by S. ambofaciens. Indeed, the deletion of this gene affects the production of the spiramycin and stambomycin macrolide antibiotics and of the grey spore pigment, all three being PKS-derived metabolites, as well as the production of the nonribosomally produced compounds, the hydroxamate siderophore coelichelin and the pyrrolamide antibiotic Congocidine. In addition, this PPTase seems to act in concert with the product of samL0372 to activate the ACP and/or PCP domains of the antimycin biosynthesis cluster which is also responsible for the production of volatile lactones.

  • Genome-guided Exploration of Streptomyces ambofaciens Secondary Metabolism
    2011
    Co-Authors: Bertrand Aigle, Sylvie Lautru, Sarka Nezbedova, Robert Bunet, Christophe Corre, Amélie Garenaux, S. Huang, Luisa Laureti, Maria Vaz Mendez, H.c. Nguyen
    Abstract:

    Members of the Streptomyces genus are among the most prolific microorganisms producing secondary metabolites with wide uses in medicine and in agriculture. Sequencing of the genome of the model Streptomyces, Streptomyces coelicolor, has highlighted an unexpected feature, i.e. that the potential of these organisms to synthesise secondary metabolites has been largely underestimated. They indeed possess many more gene clusters encoding natural product-like biosynthetic pathways than there are known natural products. Similar observations have since been made for other bacterial or fungal genomes. Thus, it became clear that microbial secondary metabolism had been seriously underestimated and that genome-based approaches were very promising for the search of new bioactive compounds. Here, we present an overview of the secondary metabolite biosynthetic potential of Streptomyces ambofaciens, a species known for decades as producer of the macrolide spiramycin and the pyrrolamide Congocidine. Interestingly, genome analysis has revealed that despite of the close phylogenetic relatedness between S. coelicolor and S. ambofaciens, most of its secondary metabolite gene clusters are species-specific.

  • Characterization and manipulation of the pathway-specific late regulator AlpW reveals Streptomyces ambofaciens as a new producer of kinamycins
    Journal of Bacteriology, 2011
    Co-Authors: Robert Bunet, Pierre Leblond, Maria Vaz Mendez, L. Song, C. Corre, L. Hotel, N. Rouhier, X. Framboisier, G.l. Challis, Bertrand Aigle
    Abstract:

    The genome sequence of Streptomyces ambofaciens, a species known to produce the Congocidine and spiramycin antibiotics, has revealed the presence of numerous gene clusters predicted to be involved in the biosynthesis of secondary metabolites. Among them, the type II polyketide synthase-encoding alp cluster was shown to be responsible for the biosynthesis of a compound with antibacterial activity. Here, by means of a deregulation approach, we gained access to workable amounts of the antibiotics for structure elucidation. These compounds, previously designated as alpomycin, were shown to be known members of kinamycin family of antibiotics. Indeed, a mutant lacking AlpW, a member of the TetR regulator family, was shown to constitutively produce kinamycins. Comparative transcriptional analyses showed that expression of alpV, the essential regulator gene required for activation of the biosynthetic genes, is strongly maintained during the stationary growth phase in the alpW mutant, a stage at which alpV transcripts and thereby transcripts of the biosynthetic genes normally drop off. Recombinant AlpW displayed DNA binding activity toward specific motifs in the promoter region of its own gene and that of alpV and alpZ. These recognition sequences are also targets for AlpZ, the -butyrolactone-like receptor involved in the regulation of the alp cluster. However, unlike that of AlpZ, the AlpW DNA-binding ability seemed to be insensitive to the signaling molecules controlling antibiotic biosynthesis. Together, the results presented in this study reveal S. ambofaciens to be a new producer of kinamycins and AlpW to be a key late repressor of the cellular control of kinamycin biosynthesis.

Maud Juguet - One of the best experts on this subject based on the ideXlab platform.

  • Natural Combinatorial Biosynthesis Involving Two Clusters for the Synthesis of Three Pyrrolamides in Streptomyces netropsis.
    ACS Chemical Biology, 2015
    Co-Authors: Audrey Vingadassalon, Florence Lorieux, Maud Juguet, Géraldine Le Goff, Claude Gerbaud, Jean-luc Pernodet, Sylvie Lautru
    Abstract:

    The pyrrolamides constitute a small family of secondary metabolites that are known for their ability to bind noncovalently to the DNA minor groove with some sequence specificity. To date, only a single pyrrolamide biosynthetic gene cluster has been reported, directing the synthesis of Congocidine (netropsin) in Streptomyces ambofaciens. In this study, we improve our understanding of pyrrolamide biosynthesis through the identification and characterization of the gene cluster responsible for the production of distamycin in Streptomyces netropsis DSM40846. We discover that the strain produces two other pyrrolamides, the well-characterized Congocidine and a Congocidine/distamycin hybrid that we named disgocidine. S. netropsis DSM40846 genome analysis led to the identification of two distinct pyrrolamide-like biosynthetic gene clusters. We show here that these two clusters are reciprocally dependent for the production of the three pyrrolamide molecules. Furthermore, based on detailed functional analysis of these clusters, we propose a biosynthetic route to Congocidine and distamycin and an updated model for pyrrolamide assembly. The synthesis of disgocidine, the distamycin/Congocidine hybrid, appears to constitute the first example of "natural combinatorial biosynthesis" between two related biosynthetic pathways. Finally, we analyze the genomic context of the two biosynthetic gene clusters and suggest that the presently interdependent clusters result from the coevolution of two ancestral independent pyrrolamide gene clusters.

  • Natural Combinatorial Biosynthesis Involving Two Clusters for the Synthesis of Three Pyrrolamides in Streptomyces netropsis
    2015
    Co-Authors: Audrey Vingadassalon, Florence Lorieux, Maud Juguet, Claude Gerbaud, Jean-luc Pernodet, Géraldine Le Goff, Sylvie Lautru
    Abstract:

    The pyrrolamides constitute a small family of secondary metabolites that are known for their ability to bind noncovalently to the DNA minor groove with some sequence specificity. To date, only a single pyrrolamide biosynthetic gene cluster has been reported, directing the synthesis of Congocidine (netropsin) in Streptomyces ambofaciens. In this study, we improve our understanding of pyrrolamide biosynthesis through the identification and characterization of the gene cluster responsible for the production of distamycin in Streptomyces netropsis DSM40846. We discover that the strain produces two other pyrrolamides, the well-characterized Congocidine and a Congocidine/distamycin hybrid that we named disgocidine. S. netropsis DSM40846 genome analysis led to the identification of two distinct pyrrolamide-like biosynthetic gene clusters. We show here that these two clusters are reciprocally dependent for the production of the three pyrrolamide molecules. Furthermore, based on detailed functional analysis of these clusters, we propose a biosynthetic route to Congocidine and distamycin and an updated model for pyrrolamide assembly. The synthesis of disgocidine, the distamycin/Congocidine hybrid, appears to constitute the first example of “natural combinatorial biosynthesis” between two related biosynthetic pathways. Finally, we analyze the genomic context of the two biosynthetic gene clusters and suggest that the presently interdependent clusters result from the coevolution of two ancestral independent pyrrolamide gene clusters

  • an iterative nonribosomal peptide synthetase assembles the pyrrole amide antibiotic Congocidine in streptomyces ambofaciens
    Chemistry & Biology, 2009
    Co-Authors: Maud Juguet, Pierre Leblond, Sylvie Lautru, Francoisxavier Francou, Sarka Nezbedova, Muriel Gondry, Jean-luc Pernodet
    Abstract:

    Congocidine (netropsin) is a pyrrole-amide (oligopyrrole, oligopeptide) antibiotic produced by Streptomyces ambofaciens. We have identified, in the right terminal region of the S. ambofaciens chromosome, the gene cluster that directs Congocidine biosynthesis. Heterologous expression of the cluster and in-frame deletions of 8 of the 22 genes confirm the involvement of this cluster in Congocidine biosynthesis. Nine genes can be assigned specific functions in regulation, resistance, or Congocidine assembly. In contrast, the biosynthetic origin of the precursors cannot be easily inferred from in silico analyses. Congocidine is assembled by a nonribosomal peptide synthetase (NRPS) constituted of a free-standing module and several single-domain proteins encoded by four genes. The iterative use of its unique adenylation domain, the utilization of guanidinoacetyl-CoA as a substrate by a condensation domain, and the control of 4-aminopyrrole-2-carboxylate polymerization constitute the most original features of this NRPS.