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Endophytic Fungus of Achyrocline satureioides: Molecular Identification, Chemical Characterization, and Cytotoxic Evaluation of its Metabolites in Human Melanoma cell line

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Abstract

Endophytic fungi are important sources of anticancer compounds. An endophytic fungus was isolated from the medicinal plant Achyrocline satureioides, and molecularly identified as Biscogniauxia sp. (family Xylariaceae) based on partial nucleotide sequences of the internal transcribed spacer genomic region (GenBank Accession No. ON257911). The chemical characterization and cytotoxic properties of secondary metabolites produced by Biscogniauxia sp. were evaluated in a human melanoma cell line (A375). The fungus was grown in potato-dextrose liquid medium for 25 days, and the extracted compounds were subjected to solid-phase fractionation to obtain the purified FDCM fraction, for which the metabolites were elucidated via ultra-performance chromatography coupled to a mass spectrometer. In the present study, 17 secondary metabolites of Biscogniauxia sp., including nine polyketide derivatives, five terpenoids, and three isocoumarins, were putatively identified. This is the first study to report of the ability of Biscogniauxia sp. in the production of isocoumarin orthosporin; the terpenoids nigriterpene A and 10-xylariterpenoid; the polyketide derivatives daldinin C, 7’dechloro-5’-hydroxygriseofulvin, daldinone D, Sch-642305, curtachalasin A, cytochalasin E, epoxycytochalasins Z8, Z8 isomer, and Z17. Furthermore, this study has reported the biosynthesis of Sch-642305 by a Xylariaceae fungus for the first time. FDCM significantly reduced the viability and proliferation of human melanoma cells at half-maximal inhibitory concentrations ​​of 10.34 and 6.89 µg/mL, respectively, and induced late apoptosis/necrosis and cell cycle arrest in G2/M phase after 72 h of treatment. Given its ability to produce unique metabolites with promising cytotoxic effects, Biscogniauxia sp. of A. satureioides may be a reservoir of compounds with important therapeutic applications.

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Funding

This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS), and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brasil; finance code: 001). R.M.S is a recipient of the CNPq fellowship (310472/2021-0).

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Authors and Affiliations

  1. Programa de Pós-Graduação em Bioquímica e Bioprospecção - Laboratório de Neuroquímica, Inflamação e Câncer, Campus Capão do Leão s/n, Universidade Federal de Pelotas, Pelotas, RS, Brazil

    Nathalia Stark Pedra & Roselia Maria Spanevello

  2. Embrapa Agroindústria Tropical, Fortaleza, CE, Brazil

    Kirley Marques Canuto, Ana Sheila de Queiroz Souza & Paulo Riceli Vasconcelos Ribeiro

  3. Programa de Pós-Graduação em Bioquímica e Bioprospecção - Laboratório de Biomarcadores, Universidade Federal de Pelotas, Pelotas, RS, Brazil

    Natália Pontes Bona

  4. Centro de Ciências Agrárias/Fitossanidade, Universidade Federal de Alagoas, Rio Largo, Alagoas, Brazil

    Roberto Ramos-Sobrinho

  5. Programa de Pós-Graduação em Biociências, Universidade Federal de Ciências da Saúde de Porto Alegre, Porto Alegre, RS, Brazil

    Priscila Oliveira de Souza & Elizandra Braganhol

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  1. Nathalia Stark Pedra
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Contributions

All the authors contributed to the conception and design of this study. Material preparation, data collection, and fungal analysis were performed by Nathalia Pedra, Kirley Canuto, Ana Souza, and Paulo Ribeiro. Material preparation, data collection, and cells analyses were performed by Nathalia Pedra, Natália Bona, Priscila Souza, Roselia Spanevello, and Elizandra Braganhol. The first draft of the manuscript was written by Nathalia Pedra, and all the authors commented on the previous versions of the manuscript. All authors have read and approved the final manuscript.

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Correspondence to Nathalia Stark Pedra or Roselia Maria Spanevello.

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12010_2023_4328_MOESM1_ESM.docx

The phylogenetic relationship based on the ITS sequences of Biscogniauxia sp. is available as Supplemental Information (Fig. S1). The spectra and proposed tandem mass spectrometry (MS/MS) fragmentation mechanisms of compounds 4, 8, 13, 14, 18, 22, 25, 26, and 28 (Fig. S2-S10), and the high resolution mass spectra of the compounds 6, 7, 10, 12, 15, 17, 23 and 27 (Fig. S11-S18) are provided in the Supporting Information.

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Pedra, N.S., Canuto, K.M., de Queiroz Souza, A.S. et al. Endophytic Fungus of Achyrocline satureioides: Molecular Identification, Chemical Characterization, and Cytotoxic Evaluation of its Metabolites in Human Melanoma cell line. Appl Biochem Biotechnol 195, 4011–4035 (2023). https://doi.org/10.1007/s12010-023-04328-w

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