Ferramentas quimiométricas para avaliação do perfil químico e biológico do fungo endofítico Hypomontagnella monticulosa isolado da alga vermelha Dichotomaria marginata
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Data
2023-01-13
Autores
Moura, Lucas Henrique Silva
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Universidade Estadual Paulista (Unesp)
Resumo
Estudos relacionados ao bioma marinho, evidenciam uma rica fonte de substâncias bioativas provenientes de esponjas, moluscos e macroalgas, onde as algas vermelhas (Rhodophyceae) são consideradas uma das principais fontes produtoras de substâncias bioativas dentre as macroalgas. Fungos endofíticos de origem marinha, tornaram-se uma alternativa para a bioprospecção de novos metabólitos durante os últimos anos. Estudos anteriores, realizados em nosso grupo de pesquisa com a alga vermelha Dichotomaria marginata, permitiram isolar a linhagem de Hypomontagnella monticulosa, cujo extrato de sua incubação apresentou atividade contra inibição de crescimento bacteriano e atividade inibitória de protease. A fim de se aprofundar mais sobre o fungo, utilizou-se da quimiometria e redes moleculares (GNPS), no intuito de se observar a variação metabólica durante seu crescimento fúngico e, principalmente a variação do perfil químico quando inserido diferentes parâmetros durante seu cultivo, utilizando-se das técnicas de HPLC-DAD, LC MS, GC-MS e UPLC-ESI-QqTOF-MS/MS, para análise do perfil químico e elucidação estrutural pelas técnicas de Infravermelho e RMN de 1H e 13C. Foi possível isolar uma lactona tetracíclica tetraidroxilada 2,8,10,10-tetraidroxi-6,10- diidro-5H-fluoreno[4,5-bcd]oxepin-5-ona, também denominada como Hypoxepinona A, uma substância ainda não relatada na literatura. Verificou-se a atividade antibacteriana, anticolinesterásica, fungicida e larvicida frente ao Aedes aegypti, onde os extratos e substância isolada não apresentaram atividade significativa. Em contrapartida, os extratos apresentaram alta citotoxicidade contra linhagem de carcinoma colorretal humano (HCT116) e câncer de mama humano (MCF7), a uma concentração de 50 μg.mL-1 . A utilização de ferramentas quimiométrias auxiliou na observação da variação metabólica do fungo nos diferentes períodos de crescimento, bem como a variação quando inseridas variáveis externas durante seu crescimento promovendo uma maior quimiodiversidade, contribuindo para estudos de bioprospecção do ambiente marinho, bem como para uma melhor compreensão do metabolismo do fungo H. monticulosa.
Studies related to the marine biome show a rich source of bioactive substances from sponges, mollusks and macroalgae, where red algae (Rhodophyceae) are considered one of the main sources of bioactive substances among macroalgae. Endophytic fungi of marine origin have become an alternative for the bioprospection of new metabolites during the last years. Previous studies, carried out in our research group with the red alga Dichotomaria marginata, allowed the isolation of the Hypomontagnella monticulosa strain, whose incubation extract showed activity against bacterial growth inhibition and protease inhibitory activity. In order to go deeper into the fungus, chemometrics and molecular networks (GNPS) were used, in order to observe the metabolic variation during its fungal growth and, mainly, the variation of the chemical profile when different parameters were inserted during its cultivation, using the techniques of HPLC DAD, LC-MS, GC-MS and UPLC-ESI-QqTOF-MS/MS, for chemical profile analysis and structural elucidation by Infrared and 1H and 13C NMR techniques. It was possible to isolate a tetrahydroxylated tetracyclic lactone 2,8,10,10- tetrahydroxy-6,10-dihydro-5H-fluoreno[4,5-bcd]oxepin-5-one, also known as Hypoxepinone A, a substance not yet reported in the literature. The antibacterial, anticholinesterase, fungicidal and larvicidal activity against Aedes aegypti was verified, where the extracts and isolated substance did not show significant activity. In contrast, the extracts showed high cytotoxicity against human colorectal carcinoma (HCT116) and human breast cancer (MCF7) cell lines, at a concentration of 50 μg.mL-1 . The use of chemometric tools helped to observe the metabolic variation of the fungus in different periods of growth, as well as the variation when external variables were inserted during its growth, promoting greater chemodiversity, contributing to bioprospecting studies of the marine environment, as well as to a better understanding of the metabolism of the fungus H. monticulosa.
Studies related to the marine biome show a rich source of bioactive substances from sponges, mollusks and macroalgae, where red algae (Rhodophyceae) are considered one of the main sources of bioactive substances among macroalgae. Endophytic fungi of marine origin have become an alternative for the bioprospection of new metabolites during the last years. Previous studies, carried out in our research group with the red alga Dichotomaria marginata, allowed the isolation of the Hypomontagnella monticulosa strain, whose incubation extract showed activity against bacterial growth inhibition and protease inhibitory activity. In order to go deeper into the fungus, chemometrics and molecular networks (GNPS) were used, in order to observe the metabolic variation during its fungal growth and, mainly, the variation of the chemical profile when different parameters were inserted during its cultivation, using the techniques of HPLC DAD, LC-MS, GC-MS and UPLC-ESI-QqTOF-MS/MS, for chemical profile analysis and structural elucidation by Infrared and 1H and 13C NMR techniques. It was possible to isolate a tetrahydroxylated tetracyclic lactone 2,8,10,10- tetrahydroxy-6,10-dihydro-5H-fluoreno[4,5-bcd]oxepin-5-one, also known as Hypoxepinone A, a substance not yet reported in the literature. The antibacterial, anticholinesterase, fungicidal and larvicidal activity against Aedes aegypti was verified, where the extracts and isolated substance did not show significant activity. In contrast, the extracts showed high cytotoxicity against human colorectal carcinoma (HCT116) and human breast cancer (MCF7) cell lines, at a concentration of 50 μg.mL-1 . The use of chemometric tools helped to observe the metabolic variation of the fungus in different periods of growth, as well as the variation when external variables were inserted during its growth, promoting greater chemodiversity, contributing to bioprospecting studies of the marine environment, as well as to a better understanding of the metabolism of the fungus H. monticulosa.
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Palavras-chave
Citotoxicidade, Microorganismos, Quimiometria, Fungos endofíticos, Produtos naturais