Identification and dereplication of endophytic Colletotrichum strains by MALDI TOF mass spectrometry and molecular networking

Barthélemy, Morgane; Guérineau, Vincent; Genta‐Jouve, Grégory; Roy, Mélanie; Chave, Jérôme; Guillot, Régis; Pellissier, Léonie; Wolfender, Jean‐Luc; Stien, Didier; Éparvier, Véronique; Touboul, David

doi:10.1038/s41598-020-74852-w

Cited by 14 publications

(11 citation statements)

References 65 publications

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“…Previous work on this plant model has shown, in another collection of endophytes, that it contained some cytotoxic and antimicrobial strains in which some bioactive compounds have been identified (Barthélemy et al, 2019;Donald et al, 2019;Barthélemy et al, 2020). In the present work, a collection of fungi isolated as cultivable endophyte strains from the leaves of A. sciophilum were cultured, extracted, and screened for their effects on cell proliferation and Wnt pathway inhibition.…”

Section: Introductionmentioning

confidence: 87%

Isolation and Identification of Isocoumarin Derivatives With Specific Inhibitory Activity Against Wnt Pathway and Metabolome Characterization of Lasiodiplodia venezuelensis

et al. 2021

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The Wnt signaling pathway controls multiple events during embryonic development of multicellular animals and is carcinogenic when aberrantly activated in adults. Breast cancers are dependent on Wnt pathway overactivation mostly through dysregulation of pathway component protein expression, which necessitates the search for therapeutically relevant compounds targeting them. Highly diverse microorganisms as endophytes represent an underexplored field in the therapeutic natural products research. In the present work, the objective was to explore the chemical diversity and presence of selective Wnt inhibitors within a unique collection of fungi isolated as foliar endophytes from the long-lived tropical palm Astrocaryum sciophilum. The fungi were cultured, extracted with ethyl acetate, and screened for their effects on the Wnt pathway and cell proliferation. The endophytic strain Lasiodiplodia venezuelensis was prioritized for scaled-up fractionation based on its selective activity. Application of geometric transfer from analytical HPLC conditions to semi-preparative scale and use of dry load sample introduction enabled the isolation of 15 pure compounds in a single step. Among the molecules identified, five are original natural products described for the first time, and six are new to this species. An active fraction obtained by semi-preparative HPLC was re-purified by UHPLC-PDA using a 1.7 µm phenyl column. 75 injections of 8 µg were necessary to obtain sufficient amounts of each compound for structure elucidation and bioassays. Using this original approach, in addition to the two major compounds, a third minor compound identified as (R)-(-)-5-hydroxymellein (18) was obtained, which was found to be responsible for the significant Wnt inhibition activity recorded. Further studies of this compound and its structural analogs showed that only 18 acts in a highly specific manner, with no acute cytotoxicity. This compound is notably selective for upstream components of the Wnt pathway and is able to inhibit the proliferation of three triple negative breast cancer cell lines. In addition to the discovery of Wnt inhibitors of interest, this study contributes to better characterize the biosynthetic potential of L. venezuelensis.

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Section: Introductionmentioning

confidence: 87%

Isolation and Identification of Isocoumarin Derivatives With Specific Inhibitory Activity Against Wnt Pathway and Metabolome Characterization of Lasiodiplodia venezuelensis

et al. 2021

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“…We have successfully identified Rhibizobial species on the inhouse extended Bruker database. 59 Furthermore, Forty-two Endophytic fungal ( Colletotrichum ) isolates from Astrocaryum sciophilum , ommon palm trees of amazon forest were verified by Bruker MALDI MS platform coupled with Biotyper 97 and the analysis grouped the isolates into 18 distinctive groups and thus the authors had elucidated the strain variability of the variousColletotrichum strains using MALDI MS tool.…”

Section: Maldi Ms-based Identification Of Plant-associated Microbesmentioning

confidence: 99%

A systematic assessment of matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) application for rapid identification of pathogenic microbes that affect food crops: delivered and future deliverables

et al. 2023

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“…The majority of studied microorganisms are from French Guiana and were studied in previous research projects to explore their metabolome. Briefly, our environmental strain collection includes endophytes, insect-associated microorganisms and entomo-or phytopathogens [33][34][35][36]. All microorganisms were isolated on Potato Dextrose Agar (PDA, Condalab, Madrid, Spain) medium at 28 • C and stored at −80 • C in a solution of water and glycerol (20:80).…”

Section: Microorganisms and Culturesmentioning

confidence: 99%

Classification of Environmental Strains from Order to Genus Levels Using Lipid and Protein MALDI-ToF Fingerprintings and Chemotaxonomic Network Analysis

et al. 2022

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During the last two decades, MALDI-ToF mass spectrometry has become an efficient and widely-used tool for identifying clinical isolates. However, its use for classification and identification of environmental microorganisms remains limited by the lack of reference spectra in current databases. In addition, the interpretation of the classical dendrogram-based data representation is more difficult when the quantity of taxa or chemotaxa is larger, which implies problems of reproducibility between users. Here, we propose a workflow including a concurrent standardized protein and lipid extraction protocol as well as an analysis methodology using the reliable spectra comparison algorithm available in MetGem software. We first validated our method by comparing protein fingerprints of highly pathogenic bacteria from the Robert Koch Institute (RKI) open database and then implemented protein fingerprints of environmental isolates from French Guiana. We then applied our workflow for the classification of a set of protein and lipid fingerprints from environmental microorganisms and compared our results to classical genetic identifications using 16S and ITS region sequencing for bacteria and fungi, respectively. We demonstrated that our protocol allowed general classification at the order and genus level for bacteria whereas only the Botryosphaeriales order can be finely classified for fungi.

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Identification and dereplication of endophytic Colletotrichum strains by MALDI TOF mass spectrometry and molecular networking

Cited by 14 publications

References 65 publications

Isolation and Identification of Isocoumarin Derivatives With Specific Inhibitory Activity Against Wnt Pathway and Metabolome Characterization of Lasiodiplodia venezuelensis

Isolation and Identification of Isocoumarin Derivatives With Specific Inhibitory Activity Against Wnt Pathway and Metabolome Characterization of Lasiodiplodia venezuelensis

A systematic assessment of matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) application for rapid identification of pathogenic microbes that affect food crops: delivered and future deliverables

Classification of Environmental Strains from Order to Genus Levels Using Lipid and Protein MALDI-ToF Fingerprintings and Chemotaxonomic Network Analysis

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