Co-cultivo dos fungos endofíticos Botryosphaeria parva e Saccharicola sp.: avaliação da produção metabólica
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Data
2017-03-10
Autores
Costa, Mayra Fonseca [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
Historicamente, a maioria dos novos medicamentos tem sido gerada a partir de produtos naturais, os quais são definidos como metabólitos secundários de microrganismos, plantas ou animais. Atualmente o elemento-chave para a investigação de produtos naturais é o acesso a novas fontes biológicas. Nos últimos anos, um grande número de substâncias biologicamente ativas estão sendo encontrados em diversos microrganismos, as quais são produzidas pela ativação de vias biossintéticas onde vários métodos têm sido estudados para a ativação destas. Um dos métodos utilizados com este objetivo é a cultura mista de microrganismos. Metabólitos secundários de fungos têm atraído um interesse considerável, uma vez que muitos deles apresentam estruturas únicas e atividades biológicas relevantes. Uma importante classe de fungos que vem se destacando na produção de novos compostos bioativos são os fungos endofíticos. Diante disso, visando contribuir com o desenvolvimento de novos produtos naturais bioativos e entender como ocorrem as interações entre os microrganismos, além de avaliar seus potenciais biológicos, é que nesse trabalho apresentamos a realização da co-cultura de dois endófitos: Botryosphaeria parva e Saccharicola sp isolados da espécie vegetal Eugenia jambolana. Estes endófitos foram cultivados em escala reduzida (isoladamente) e escala ampliada (co-cultivo) em Czapek (300 mL) para obtenção dos extratos brutos AcOEt. Os extratos brutos AcOEt obtidos foram submetidos a análises químicas (cromatografia em camada delgada comparativa, cromatografia líquida de alta eficiência e ressonância magnética nuclear de hidrogênio) e biológicas (ensaios antioxidante, antifúngico e citotóxico). Os extratos brutos apresentaram pelo menos uma atividade biológica positiva, sendo que no ensaio antifúngico, para C. sphaerospermum apenas o extrato Czapeck do co-cultivo codificado como CC-04/01 apresentou-se ativo. O co-cultivo dos fungos endofíticos Saccharicola sp. codificado como Ej-c4 e Botryosphaeria parva codificado como Ej-f1 isolado do caule e da folha de E. jambolana, respectivamente, permitiu o isolamento e elucidação de nove substâncias sendo uma inédita (2). Foram isoladas seis substâncias (1, 3, 4, 5, 6, 7, 8 e 9) produzidas pelo endófito Botryosphaeria parva (Ej-f1), e uma substância (2) produzida por Saccharicola sp. As estruturas das substâncias isoladas foram elucidadas pelo uso de técnicas espectrométricas como RMN uni e bidimensional e espectrometria de massas.
Historically, most new drugs have been generated from natural products, which are defined as secondary metabolites or byproducts of microorganisms, plants or animals. Currently, the key element for the investigation of natural products is the access to new biological sources. In recent years, a large number of biologically active substances are being reported in several microorganisms, which are produced by the activation of biosynthetic pathways where several methods have been studied for their activation. One of the methods used for this purpose is the mixed culture of microorganisms. Secondary fungal metabolites have attracted considerable interest, since many of them have unique structures and relevant biological activities. An important class of fungi that has been prominent in the production of new bioactive compounds are endophytic fungi. Therefore, in this study we present the co-culture of two endophytes: Botryosphaeria parva and Saccharicola sp. isolated from the plant species Eugenia jambolana. Our objective is to contribute to the development of new bioactive natural products and to understand how the chemical interactions of these microorganisms occur, as well as to evaluate their biological potential. These endophytes were cultivated on a reduced scale (isolated) and extended scale (co-culture) in Czapek (300 mL) to obtain the crude extracts AcOEt. The crude AcOEt extracts obtained were submitted to chemical analysis (comparative thin layer chromatography, high performance liquid chromatography and nuclear magnetic resonance of hydrogen) and biological tests (antioxidant, antifungal and cytotoxic assays). The crude extracts presented at least one positive biological activity, whereas in the antifungal assay, for C. sphaerospermum only the Czapeck extract of the cocultivation codified as CC-04/01 was active. The co-cultivation of the endophytic fungi Saccharicola sp. Coded as Ej-c4 and Botryosphaeria parva coded as Ej-f1 isolated from the stem and leaf of E. jambolana, respectively, allowed the isolation and elucidation of nine substances and one of them wasunprecedented (2). Six substances (1, 3, 4, 5, 6, 7, 8 and 9) produced by the endophyte Botryosphaeria parva (Ej-f1), and a substance (2) produced by Saccharicola sp. The structures of the isolated substances were elucidated by the use of spectrometric techniques such as uni and bidimensional NMR and mass spectrometry.
Historically, most new drugs have been generated from natural products, which are defined as secondary metabolites or byproducts of microorganisms, plants or animals. Currently, the key element for the investigation of natural products is the access to new biological sources. In recent years, a large number of biologically active substances are being reported in several microorganisms, which are produced by the activation of biosynthetic pathways where several methods have been studied for their activation. One of the methods used for this purpose is the mixed culture of microorganisms. Secondary fungal metabolites have attracted considerable interest, since many of them have unique structures and relevant biological activities. An important class of fungi that has been prominent in the production of new bioactive compounds are endophytic fungi. Therefore, in this study we present the co-culture of two endophytes: Botryosphaeria parva and Saccharicola sp. isolated from the plant species Eugenia jambolana. Our objective is to contribute to the development of new bioactive natural products and to understand how the chemical interactions of these microorganisms occur, as well as to evaluate their biological potential. These endophytes were cultivated on a reduced scale (isolated) and extended scale (co-culture) in Czapek (300 mL) to obtain the crude extracts AcOEt. The crude AcOEt extracts obtained were submitted to chemical analysis (comparative thin layer chromatography, high performance liquid chromatography and nuclear magnetic resonance of hydrogen) and biological tests (antioxidant, antifungal and cytotoxic assays). The crude extracts presented at least one positive biological activity, whereas in the antifungal assay, for C. sphaerospermum only the Czapeck extract of the cocultivation codified as CC-04/01 was active. The co-cultivation of the endophytic fungi Saccharicola sp. Coded as Ej-c4 and Botryosphaeria parva coded as Ej-f1 isolated from the stem and leaf of E. jambolana, respectively, allowed the isolation and elucidation of nine substances and one of them wasunprecedented (2). Six substances (1, 3, 4, 5, 6, 7, 8 and 9) produced by the endophyte Botryosphaeria parva (Ej-f1), and a substance (2) produced by Saccharicola sp. The structures of the isolated substances were elucidated by the use of spectrometric techniques such as uni and bidimensional NMR and mass spectrometry.
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Palavras-chave
Fungo endofítico, Eugenia Jambolana Lam, Botryosphaeria parva, Saccharicola sp., Co-cultivo