Stereomeric Lipopeptides from a Single Non-Ribosomal Peptide Synthetase as an Additional Source of Structural and Functional Diversification in <i>Pseudomonas</i> Lipopeptide Biosynthesis
In <i>Pseudomonas</i> lipopeptides, the D-configuration of amino acids is generated by dedicated, dual-function epimerization/condensation (E/C) domains. The increasing attention to stereochemistry in lipopeptide structure elucidation efforts has revealed multiple examples where epimeriz...
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2023-09-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/18/14302 |
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| author | Penthip Muangkaew Vic De Roo Lu Zhou Léa Girard Catherine Cesa-Luna Monica Höfte René De Mot Annemieke Madder Niels Geudens José C. Martins |
| author_facet | Penthip Muangkaew Vic De Roo Lu Zhou Léa Girard Catherine Cesa-Luna Monica Höfte René De Mot Annemieke Madder Niels Geudens José C. Martins |
| author_sort | Penthip Muangkaew |
| collection | DOAJ |
| description | In <i>Pseudomonas</i> lipopeptides, the D-configuration of amino acids is generated by dedicated, dual-function epimerization/condensation (E/C) domains. The increasing attention to stereochemistry in lipopeptide structure elucidation efforts has revealed multiple examples where epimerization does not occur, even though an E/C-type domain is present. While the origin of the idle epimerization in those E/C-domains remains elusive, epimerization activity has so far shown a binary profile: it is either ‘on’ (active) or ‘off’ (inactive). Here, we report the unprecedented observation of an E/C-domain that acts ‘on and off’, giving rise to the production of two diastereoisomeric lipopeptides by a single non-ribosomal peptide synthetase system. Using dereplication based on solid-phase peptide synthesis and NMR fingerprinting, we first show that the two cyclic lipopeptides produced by <i>Pseudomonas entomophila</i> COR5 correspond to entolysin A and B originally described for <i>P. entomophila</i> L48. Next, we prove that both are diastereoisomeric homologues differing only in the configuration of a single amino acid. This configurational variability is maintained in multiple <i>Pseudomonas</i> strains and typically occurs in a 3:2 ratio. Bioinformatic analysis reveals a possible correlation with the composition of the flanking sequence of the N-terminal secondary histidine motif characteristic for dual-function E/C-type domains. In permeabilization assays, using propidium iodide entolysin B has a higher antifungal activity compared to entolysin A against <i>Botrytis cinerea</i> and <i>Pyricularia oryzae</i> spores. The fact that configurational homologues are produced by the same NRPS system in a <i>Pseudomonas</i> strain adds a new level of structural and functional diversification to those already known from substrate flexibility during the recruitment of the amino acids and fatty acids and underscores the importance of complete stereochemical elucidation of non-ribosomal lipopeptide structures. |
| first_indexed | 2024-03-10T22:39:40Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T22:39:40Z |
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| spelling | doaj.art-d68069bd9b27454ba49d3a2ef8e152fb2023-11-19T11:10:46ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-09-0124181430210.3390/ijms241814302Stereomeric Lipopeptides from a Single Non-Ribosomal Peptide Synthetase as an Additional Source of Structural and Functional Diversification in <i>Pseudomonas</i> Lipopeptide BiosynthesisPenthip Muangkaew0Vic De Roo1Lu Zhou2Léa Girard3Catherine Cesa-Luna4Monica Höfte5René De Mot6Annemieke Madder7Niels Geudens8José C. Martins9Organic and Biomimetic Chemistry Research Group, Department of Organic and Macromolecular Chemistry, Faculty of Science, Ghent University, B-9000 Ghent, BelgiumOrganic and Biomimetic Chemistry Research Group, Department of Organic and Macromolecular Chemistry, Faculty of Science, Ghent University, B-9000 Ghent, BelgiumLaboratory of Phytopathology, Department of Plants and Crops, Faculty of Bioscience Engineering, Ghent University, B-9000 Ghent, BelgiumCentre of Microbial and Plant Genetics, Faculty of Bioscience Engineering, Katholieke Universiteit Leuven, B-3001 Heverlee, BelgiumCentre of Microbial and Plant Genetics, Faculty of Bioscience Engineering, Katholieke Universiteit Leuven, B-3001 Heverlee, BelgiumLaboratory of Phytopathology, Department of Plants and Crops, Faculty of Bioscience Engineering, Ghent University, B-9000 Ghent, BelgiumCentre of Microbial and Plant Genetics, Faculty of Bioscience Engineering, Katholieke Universiteit Leuven, B-3001 Heverlee, BelgiumOrganic and Biomimetic Chemistry Research Group, Department of Organic and Macromolecular Chemistry, Faculty of Science, Ghent University, B-9000 Ghent, BelgiumNMR and Structure Analysis Unit, Department of Organic and Macromolecular Chemistry, Faculty of Science, Ghent University, B-9000 Ghent, BelgiumNMR and Structure Analysis Unit, Department of Organic and Macromolecular Chemistry, Faculty of Science, Ghent University, B-9000 Ghent, BelgiumIn <i>Pseudomonas</i> lipopeptides, the D-configuration of amino acids is generated by dedicated, dual-function epimerization/condensation (E/C) domains. The increasing attention to stereochemistry in lipopeptide structure elucidation efforts has revealed multiple examples where epimerization does not occur, even though an E/C-type domain is present. While the origin of the idle epimerization in those E/C-domains remains elusive, epimerization activity has so far shown a binary profile: it is either ‘on’ (active) or ‘off’ (inactive). Here, we report the unprecedented observation of an E/C-domain that acts ‘on and off’, giving rise to the production of two diastereoisomeric lipopeptides by a single non-ribosomal peptide synthetase system. Using dereplication based on solid-phase peptide synthesis and NMR fingerprinting, we first show that the two cyclic lipopeptides produced by <i>Pseudomonas entomophila</i> COR5 correspond to entolysin A and B originally described for <i>P. entomophila</i> L48. Next, we prove that both are diastereoisomeric homologues differing only in the configuration of a single amino acid. This configurational variability is maintained in multiple <i>Pseudomonas</i> strains and typically occurs in a 3:2 ratio. Bioinformatic analysis reveals a possible correlation with the composition of the flanking sequence of the N-terminal secondary histidine motif characteristic for dual-function E/C-type domains. In permeabilization assays, using propidium iodide entolysin B has a higher antifungal activity compared to entolysin A against <i>Botrytis cinerea</i> and <i>Pyricularia oryzae</i> spores. The fact that configurational homologues are produced by the same NRPS system in a <i>Pseudomonas</i> strain adds a new level of structural and functional diversification to those already known from substrate flexibility during the recruitment of the amino acids and fatty acids and underscores the importance of complete stereochemical elucidation of non-ribosomal lipopeptide structures.https://www.mdpi.com/1422-0067/24/18/14302<i>Pseudomonas entomophila</i>cyclic lipodepsipeptideentolysinstereochemistrynon-ribosomal peptide synthetaseE/C-type domain |
| spellingShingle | Penthip Muangkaew Vic De Roo Lu Zhou Léa Girard Catherine Cesa-Luna Monica Höfte René De Mot Annemieke Madder Niels Geudens José C. Martins Stereomeric Lipopeptides from a Single Non-Ribosomal Peptide Synthetase as an Additional Source of Structural and Functional Diversification in <i>Pseudomonas</i> Lipopeptide Biosynthesis International Journal of Molecular Sciences <i>Pseudomonas entomophila</i> cyclic lipodepsipeptide entolysin stereochemistry non-ribosomal peptide synthetase E/C-type domain |
| title | Stereomeric Lipopeptides from a Single Non-Ribosomal Peptide Synthetase as an Additional Source of Structural and Functional Diversification in <i>Pseudomonas</i> Lipopeptide Biosynthesis |
| title_full | Stereomeric Lipopeptides from a Single Non-Ribosomal Peptide Synthetase as an Additional Source of Structural and Functional Diversification in <i>Pseudomonas</i> Lipopeptide Biosynthesis |
| title_fullStr | Stereomeric Lipopeptides from a Single Non-Ribosomal Peptide Synthetase as an Additional Source of Structural and Functional Diversification in <i>Pseudomonas</i> Lipopeptide Biosynthesis |
| title_full_unstemmed | Stereomeric Lipopeptides from a Single Non-Ribosomal Peptide Synthetase as an Additional Source of Structural and Functional Diversification in <i>Pseudomonas</i> Lipopeptide Biosynthesis |
| title_short | Stereomeric Lipopeptides from a Single Non-Ribosomal Peptide Synthetase as an Additional Source of Structural and Functional Diversification in <i>Pseudomonas</i> Lipopeptide Biosynthesis |
| title_sort | stereomeric lipopeptides from a single non ribosomal peptide synthetase as an additional source of structural and functional diversification in i pseudomonas i lipopeptide biosynthesis |
| topic | <i>Pseudomonas entomophila</i> cyclic lipodepsipeptide entolysin stereochemistry non-ribosomal peptide synthetase E/C-type domain |
| url | https://www.mdpi.com/1422-0067/24/18/14302 |
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