Prospecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae Mez

Detalhes bibliográficos
Autor(a) principal: Fernanda Motta Ribeiro da Silva
Data de Publicação: 2023
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFMS
Texto Completo: https://repositorio.ufms.br/handle/123456789/5906
Resumo: Fungi biosynthesize metabolites with high diversity and structural complexity that are potentially bioactive. The rocky outcrops occur in the region of the western edge of the Pantanal Sul-mato-grossense, Brazil, which are inhospitable places, their plant species may be adapted for these locals and the symbiosis with microorganisms can help their defense system. The present work aimed to bioprospect of bioactive metabolites from the endophytic fungi of Bromelia balansae Mez, collected in Pantanal RA. Sixty-one endophytic fungi were isolated and ten were molecularly identified, among them the genera: Aspergillus, Penicillium, Diaporthe, Daldinia and Neodeightonia. The isolates were grown in potato-dextrose agar medium (BDA) and in potato dextrose broth. The organic extracts obtained from these cultures were evaluated in vitro bioassays of antimicrobial, antibiofilm, antileishmania, and cytotoxic activity. Of these, 25 endophytes showed antimicrobial activity against Staphylococcus aureus with MIC (minimum inhibitory concentration) ≤ 500 µg/mL, and 12 strains showed MIC ≤ 125 µg/mL. None of the microorganisms isolated was promising against Pseudomonas aeruginosa. In the Leishmania infantum assay, four strains had an infection rate lower than 15% compared to the control. In the cytotoxicity evaluation against leukemic lines, Kasumi and KG-1, five isolates inhibited more than 90% of cell growth at the concentration of 100 µg/mL. The isolate 5MP2F4, identified as Penicillium sp., showed MIC of 32 µg/mL against S. aureus, and therefore, it was selected for a study using the OSMAC strategy, antibacterial and antibiofilm screening, with application of metabolomics, statistical analysis, and molecular networking to determine potential bioactive metabolites and understanding of the chemical profile of the fungus. In this experiment, the obtained extracts showed different chemical profiles and antibacterial activity. Several metabolites were annotated, among them different braziliamides, verruculogen, fumitremorgin, penicillic acid, viridicatum toxin A, and spirohexalin. An optimization study of extraction from fungal mycelia from the isolate 5MP2F4 by accelerated solvent extraction (ASE) was also performed, and viridicatum toxin A (VRA) was purified by this methodology. VRA was evaluated for the activities anti-leishmanial, antibiofilm, and cytotoxic against leukemic cell lines, and it showed to be active in all experiments. Fungal isolates of B. balansae have expanded knowledge of the endophyte biodiversity of the Pantanal. Several substances belonging to different chemical classes were found, some were produced in high levels according to the variation in the culture medium, and the bioassays suggest that they have potential as antibiotics and cytotoxics.
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spelling 2023-05-16T19:57:41Z2023-05-16T19:57:41Z2023https://repositorio.ufms.br/handle/123456789/5906Fungi biosynthesize metabolites with high diversity and structural complexity that are potentially bioactive. The rocky outcrops occur in the region of the western edge of the Pantanal Sul-mato-grossense, Brazil, which are inhospitable places, their plant species may be adapted for these locals and the symbiosis with microorganisms can help their defense system. The present work aimed to bioprospect of bioactive metabolites from the endophytic fungi of Bromelia balansae Mez, collected in Pantanal RA. Sixty-one endophytic fungi were isolated and ten were molecularly identified, among them the genera: Aspergillus, Penicillium, Diaporthe, Daldinia and Neodeightonia. The isolates were grown in potato-dextrose agar medium (BDA) and in potato dextrose broth. The organic extracts obtained from these cultures were evaluated in vitro bioassays of antimicrobial, antibiofilm, antileishmania, and cytotoxic activity. Of these, 25 endophytes showed antimicrobial activity against Staphylococcus aureus with MIC (minimum inhibitory concentration) ≤ 500 µg/mL, and 12 strains showed MIC ≤ 125 µg/mL. None of the microorganisms isolated was promising against Pseudomonas aeruginosa. In the Leishmania infantum assay, four strains had an infection rate lower than 15% compared to the control. In the cytotoxicity evaluation against leukemic lines, Kasumi and KG-1, five isolates inhibited more than 90% of cell growth at the concentration of 100 µg/mL. The isolate 5MP2F4, identified as Penicillium sp., showed MIC of 32 µg/mL against S. aureus, and therefore, it was selected for a study using the OSMAC strategy, antibacterial and antibiofilm screening, with application of metabolomics, statistical analysis, and molecular networking to determine potential bioactive metabolites and understanding of the chemical profile of the fungus. In this experiment, the obtained extracts showed different chemical profiles and antibacterial activity. Several metabolites were annotated, among them different braziliamides, verruculogen, fumitremorgin, penicillic acid, viridicatum toxin A, and spirohexalin. An optimization study of extraction from fungal mycelia from the isolate 5MP2F4 by accelerated solvent extraction (ASE) was also performed, and viridicatum toxin A (VRA) was purified by this methodology. VRA was evaluated for the activities anti-leishmanial, antibiofilm, and cytotoxic against leukemic cell lines, and it showed to be active in all experiments. Fungal isolates of B. balansae have expanded knowledge of the endophyte biodiversity of the Pantanal. Several substances belonging to different chemical classes were found, some were produced in high levels according to the variation in the culture medium, and the bioassays suggest that they have potential as antibiotics and cytotoxics.Os fungos são produtores de metabólitos com elevada diversidade e complexidade estrutural e que são potencialmente bioativos. Os afloramentos rochosos (AR) que ocorrem na região da borda oeste do Pantanal Sul-mato grossense, Brasil, são locais inóspitos, por isso acredita-se que as espécies vegetais que existem neste local podem estar adaptadas e em simbiose com microrganismos que ajudam o seu sistema de defesa. O presente trabalho teve como objetivo a bioprospecção de metabólitos bioativos de fungos endofíticos de Bromelia balansae Mez, coletada em AR do Pantanal. Foram isolados 61 fungos endofíticos, sendo que dez foram identificados molecularmente, entre os quais foram identificados os gêneros: Aspergillus, Penicillium, Diaporthe, Daldinia e Neodeightonia. Os isolados foram cultivados em meio batata dextrose-ágar (BDA) e em caldo batata dextrose, os extratos orgânicos obtidos deste cultivo foram avaliados em bioensaios in vitro de atividade antimicrobiana, antibiofilme, antileishmania e citotóxica. Destes, 25 endófitos apresentaram atividade antimicrobiana frente a Staphylococcus aureus com CIM (concentração inibitória mínima) ≤ 500 µg/mL, sendo que 12 destes apresentaram CIM ≤ 125 µg/mL. Nenhum dos microrganismos isolados foi promissor frente a Pseudomonas aeruginosa. No ensaio com Leishmania infantum quatro cepas apresentaram índice de infecção menor que 15 % em relação ao controle. Na avaliação da citotoxicidade frente as linhagens leucêmicas, Kasumi e KG-1, cinco isolados inibiram mais que 90% do crescimento celular na concentração de 100 µg/mL. O isolado 5MP2F4, identificado como Penicillium sp., apresentou CIM de 32 µg/mL frente a S. aureus, e por isso, este foi selecionado para um estudo utilizando a estratégia OSMAC, triagem antibacteriana e antibiofilme, com aplicação da metabolômica, análises estatísticas e construção de rede molecular para determinação de possíveis metabólitos bioativos e compreensão do perfil químico do fungo. Neste experimento, os extratos obtidos apresentaram diferentes perfis químicos e potencial antimicrobiano. Diversos metabólitos foram anotados, dentre eles diferentes brasiliamidas, verruculogeno, fumitremorgina, ácido penicílico, viridicatum toxina A e espirohexalina. Foi realizado também um estudo de otimização da extração de micélios fúngicos do isolado 5MP2F4, utilizando extração acelerada por solvente (ASE), e a viridicatum toxina A (VRA) foi purificada a partir desta metodologia. VRA foi avaliada quanto a sua atividade anti-leishmania, antibiofilme e citotóxica frente as linhagens de células leucêmicas e seu mostrou promissora em todos os experimentos. Os isolados fúngicos de B. balansae ampliaram o conhecimento da biodiversidade de endófitos do Pantanal. Foram encontradas diversas substâncias pertencentes a diferentes classes químicas, algumas foram produzidas em teores elevados conforme a variação do meio de cultivo, e os bioensaios nos sugerem que estas possuem potencial como antibióticos e citotóxicos.Fundação Universidade Federal de Mato Grosso do SulUFMSBrasilProspecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae MezProspecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae Mezinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisDenise Brentan da SilvaFernanda Motta Ribeiro da Silvainfo:eu-repo/semantics/openAccessporreponame:Repositório Institucional da UFMSinstname:Universidade Federal de Mato Grosso do Sul (UFMS)instacron:UFMSORIGINALTESE_FernandaMottaRibeirodaSilva.pdfTESE_FernandaMottaRibeirodaSilva.pdfapplication/pdf19060436https://repositorio.ufms.br/bitstream/123456789/5906/-1/TESE_FernandaMottaRibeirodaSilva.pdfdf89b6333ac9c1cbb3f1f8976647f6dcMD5-1123456789/59062023-05-16 15:57:45.449oai:repositorio.ufms.br:123456789/5906Repositório InstitucionalPUBhttps://repositorio.ufms.br/oai/requestri.prograd@ufms.bropendoar:21242023-05-16T19:57:45Repositório Institucional da UFMS - Universidade Federal de Mato Grosso do Sul (UFMS)false
dc.title.pt_BR.fl_str_mv Prospecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae Mez
title Prospecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae Mez
spellingShingle Prospecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae Mez
Fernanda Motta Ribeiro da Silva
Prospecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae Mez
title_short Prospecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae Mez
title_full Prospecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae Mez
title_fullStr Prospecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae Mez
title_full_unstemmed Prospecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae Mez
title_sort Prospecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae Mez
author Fernanda Motta Ribeiro da Silva
author_facet Fernanda Motta Ribeiro da Silva
author_role author
dc.contributor.advisor1.fl_str_mv Denise Brentan da Silva
dc.contributor.author.fl_str_mv Fernanda Motta Ribeiro da Silva
contributor_str_mv Denise Brentan da Silva
dc.subject.por.fl_str_mv Prospecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae Mez
topic Prospecção de Produtos Naturais de Fungos Endofíticos de Bromelia balansae Mez
description Fungi biosynthesize metabolites with high diversity and structural complexity that are potentially bioactive. The rocky outcrops occur in the region of the western edge of the Pantanal Sul-mato-grossense, Brazil, which are inhospitable places, their plant species may be adapted for these locals and the symbiosis with microorganisms can help their defense system. The present work aimed to bioprospect of bioactive metabolites from the endophytic fungi of Bromelia balansae Mez, collected in Pantanal RA. Sixty-one endophytic fungi were isolated and ten were molecularly identified, among them the genera: Aspergillus, Penicillium, Diaporthe, Daldinia and Neodeightonia. The isolates were grown in potato-dextrose agar medium (BDA) and in potato dextrose broth. The organic extracts obtained from these cultures were evaluated in vitro bioassays of antimicrobial, antibiofilm, antileishmania, and cytotoxic activity. Of these, 25 endophytes showed antimicrobial activity against Staphylococcus aureus with MIC (minimum inhibitory concentration) ≤ 500 µg/mL, and 12 strains showed MIC ≤ 125 µg/mL. None of the microorganisms isolated was promising against Pseudomonas aeruginosa. In the Leishmania infantum assay, four strains had an infection rate lower than 15% compared to the control. In the cytotoxicity evaluation against leukemic lines, Kasumi and KG-1, five isolates inhibited more than 90% of cell growth at the concentration of 100 µg/mL. The isolate 5MP2F4, identified as Penicillium sp., showed MIC of 32 µg/mL against S. aureus, and therefore, it was selected for a study using the OSMAC strategy, antibacterial and antibiofilm screening, with application of metabolomics, statistical analysis, and molecular networking to determine potential bioactive metabolites and understanding of the chemical profile of the fungus. In this experiment, the obtained extracts showed different chemical profiles and antibacterial activity. Several metabolites were annotated, among them different braziliamides, verruculogen, fumitremorgin, penicillic acid, viridicatum toxin A, and spirohexalin. An optimization study of extraction from fungal mycelia from the isolate 5MP2F4 by accelerated solvent extraction (ASE) was also performed, and viridicatum toxin A (VRA) was purified by this methodology. VRA was evaluated for the activities anti-leishmanial, antibiofilm, and cytotoxic against leukemic cell lines, and it showed to be active in all experiments. Fungal isolates of B. balansae have expanded knowledge of the endophyte biodiversity of the Pantanal. Several substances belonging to different chemical classes were found, some were produced in high levels according to the variation in the culture medium, and the bioassays suggest that they have potential as antibiotics and cytotoxics.
publishDate 2023
dc.date.accessioned.fl_str_mv 2023-05-16T19:57:41Z
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publisher.none.fl_str_mv Fundação Universidade Federal de Mato Grosso do Sul
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