Fauna acompanhante: um universo químico a ser explorado
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Data
2016-06-27
Autores
Tangerina, Marcelo Marucci Pereira [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
A fauna acompanhante da pesca do camarão inclui uma série de invertebrados marinhos que são descartados por não ter valor comercial. A fim de tentar acrescentar algum valor a este material, foi analisada a composição química da estrela-do-mar Luidia senegalensis coletada na costa brasileira como consequência da aplicação da pesca de arrasto. A fim de avaliar sua composição química, foi utilizada uma combinação de extração em fase sólida (SPE) seguida de cromatografia líquida de ultra eficiência acoplada a espectrômetro de massas equipado com fonte de ionização por eletrosptray e analisador ion-trap linear (UPLCESI- IT-MSn). Luidia senegalensis contém asterosaponinas, que são esteroides glicosilados sulfatados contendo cinco e seis unidades de açúcar, além de poliidroxiesteroides. Este estudo mostrou a presença de compostos importantes e potencialmente bioativos em invertebrados associados à fauna acompanhante da pesca do camarão, usando um método rápido e eficiente. Normalmente descartada, a fauna acompanhante contém muitos invertebrados que podem hospedar uma grande variedade de gêneros de bactérias, algumas das quais com potencial de produzir produtos naturais bioativos com aplicações biotecnológicas. Assim, para utilizar um material normalmente descartado, foi explorado o potencial biotecnológico de bactérias cultiváveis de duas espécies de invertebrados abundantes na fauna acompanhante, o gastrópode Olivancillaria urceus e a estrela-do-mar Luidia senegalensis. Uma amostra de sedimento da mesma área de coleta também foi investigado. Utilizando múltiplas abordagens de isolamento 134 isolados foram obtidos a partir dos invertebrados e do sedimento. Sequenciamento parcial da subunidade de rRNA (16S) revelou que os isolados pertenciam aos filos Proteobacteria, Firmicutes e Actinobacteria, distribuídos em 28 gêneros. Vários gêneros conhecidos pela sua capacidade de produzir produtos naturais bioativos (Micromonospora, Streptomyces, Serinicoccus e Verrucosispora) foram obtidos a partir das amostras estudadas. Para avaliar as bactérias isoladas quanto à sua capacidade para produzir metabólitos bioativos todas as cepas foram fermentadas e os extratos de fermentação analisados por LC-HRMS e testados em ensaio de atividade antimicrobiana. Quatro cepas apresentaram atividade antimicrobiana contra Staphylococcus aureus resistente à meticilina (MRSA) e Staphylococcus warneri. A produção de metabólitos secundários por bactérias isoladas da fauna acompanhante também foi avaliada por uma abordagem metabolômica utilizando LC-HRMS, onde foi avaliado como as diferenças na composição dos meios de cultura podem alterar a produção de substâncias. Utilizou-se a metabolômica como uma ferramenta para investigar a produção de abyssomicinas, um agente anticâncer, e outros metabólitos secundários em três cepas do actinomiceto raro Verrucosispora maris, isoladas a partir de uma amostra de sedimento e associadas à estrela-do-mar Luidia senegalensis de Ubatuba - SP, Brasil. Nove composições diferentes de meios de cultura foram avaliadas e verificou-se que, dentre todas as cepas, somente RKMT_111 foi capaz de produzir abyssomicinas. O estudo da composição do meio de cultura revelou que a produção de abyssomicinas só foi possível em BFM-11m. Embora as três cepas pertençam à mesma espécie e são provenientes da mesma localização, é notável que cada isolado apresentou diferente capacidade de produção de metabólitos secundários. Os produtos de fermentação de Erythrobacter vulgaris foram avaliados utilizando técnicas de HPLC preparativo, LC-HRMS e RMN. A cepa foi isolada pelo método dry-stamp de uma amostra de sedimento marinho da costa de Ubatuba-SP, Brasil. Depois de sequenciamento completo do rRNA (16S) e identificação, o isolado foi fermentado em larga escala, seu caldo de fermentação extraído por solvente e os compostos purificados por HPLCMS. Análise de LC-HRMS e RMN dos compostos isolados levou à identificação de dois novos derivados do ácido cólico, ácido 3-acetil-glicocólico e o ácido 3-acetilglicodesoxicólico. As substâncias obtidas podem ter sido produzidas por biotransformação do ácido glicocólico e ácido desoxicólico, respectivamente, já presentes no meio de cultivo. Este é o primeiro relato de tais compostos e também a primeira observação de uma acilação realizada por um isolado marinho de Erythrobacter vulgaris.
The by-catch fauna of the shrimp fishery includes a number of marine invertebrates that are discarded because they do not have commercial value. In order to try to add some value to these materials, we analyzed the chemical composition of the starfish Luidia senegalensis collected in the Brazilian coast as a consequence of the trawling fishery method. In order to access their chemical composition, we used a combination of solid phase extraction (SPE) followed by ultra performance liquid chromatography coupled to electrospray ionization ion trap tandem mass spectrometry (UPLC-ESI-IT-MSn). Luidia senegalensis contains asterosaponins, which are sulphated glycosilated steroids, containing five and six sugar moieties, in addition to polyhydroxysteroids. This study helped us to support the presence of important and potentially bioactive compounds in invertebrates associated to the by-catch fauna of the shrimp fishery, using a fast and efficient method. Typically discarded, by-catch contains many invertebrates that may host a great variety of bacterial genera, some of which may produce bioactive natural products with biotechnological applications. Therefore, to utilize by-catch that is usually discarded we explored the biotechnological potential of culturable bacteria of two abundant by-catch invertebrate species, the snail Olivancillaria urceus and the sea star Luidia senegalensis. Sediment from the collection area was also investigated. Utilizing multiple isolation approaches 134 isolates were obtained from the invertebrates and sediment. Small subunit rRNA (16S) gene sequencing revealed that the isolates belonged to Proteobacteria, Firmicutes and Actinobacteria phyla and were distributed among 28 genera. Several genera known for their capacity to produce bioactive natural products (Micromonospora, Streptomyces, Serinicoccus and Verrucosispora) were retrieved from the invertebrate samples. To query the bacterial isolates for their ability to produce bioactive metabolites all strains were fermented and fermentation extracts profiled by LC-HRMS and tested for antimicrobial activity. Four strains exhibited antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus warneri. The production of secondary metabolites was assessed using a LC-HRMS-based metabolomics approach, where it was evaluated how differences in media composition can alter the production of chemical compounds. We used metabolomics as a tool to investigate the production of abyssomicins, an anticancer agent, and other secondary metabolites in three strains of the rare actinomycete Verrucosispora maris, all marine isolates from a sediment sample and associated to a starfish from the species Luidia senegalensis of Ubatuba – SP, Brazil. Nine different media compositions were evaluated and it was found that, among all strains, only RKMT_111 was capable of producing abyssomicins. The media composition study revealed that the production of abyssomicins was only achievable in BFM-11m. Although the three strains belong to the same species and the same location, it is worthwhile noticing that each isolate showed different capability for production of secondary metabolites. The products of fermentation of Erythrobacter vulgaris were evaluated using preparative HPLC, LC-HRMS and NMR techniques. Bacterial strain was isolated by drystamp method from a marine sediment sample from the coast of Ubatuba-SP, Brazil. After fully 16S rDNA sequence and identification, the marine isolate was fermented in large-scale, extracted and the compounds purified through HPLC-MS. Analysis of LC-HRMS and NMR of the isolated compounds led to the identification of two new cholic acid derivatives, 3- acetyl-glycocholic acid and 3-acetyl-glycodeoxycholic acid. Both new compounds may have been produced by the biotransformation of glycocholic acid and deoxycholic acid, respectively, already present in the cultivation medium. This is the first report of such compounds and also the first time an acylation has been observed for an Erythrobacter vulgaris marine isolate.
The by-catch fauna of the shrimp fishery includes a number of marine invertebrates that are discarded because they do not have commercial value. In order to try to add some value to these materials, we analyzed the chemical composition of the starfish Luidia senegalensis collected in the Brazilian coast as a consequence of the trawling fishery method. In order to access their chemical composition, we used a combination of solid phase extraction (SPE) followed by ultra performance liquid chromatography coupled to electrospray ionization ion trap tandem mass spectrometry (UPLC-ESI-IT-MSn). Luidia senegalensis contains asterosaponins, which are sulphated glycosilated steroids, containing five and six sugar moieties, in addition to polyhydroxysteroids. This study helped us to support the presence of important and potentially bioactive compounds in invertebrates associated to the by-catch fauna of the shrimp fishery, using a fast and efficient method. Typically discarded, by-catch contains many invertebrates that may host a great variety of bacterial genera, some of which may produce bioactive natural products with biotechnological applications. Therefore, to utilize by-catch that is usually discarded we explored the biotechnological potential of culturable bacteria of two abundant by-catch invertebrate species, the snail Olivancillaria urceus and the sea star Luidia senegalensis. Sediment from the collection area was also investigated. Utilizing multiple isolation approaches 134 isolates were obtained from the invertebrates and sediment. Small subunit rRNA (16S) gene sequencing revealed that the isolates belonged to Proteobacteria, Firmicutes and Actinobacteria phyla and were distributed among 28 genera. Several genera known for their capacity to produce bioactive natural products (Micromonospora, Streptomyces, Serinicoccus and Verrucosispora) were retrieved from the invertebrate samples. To query the bacterial isolates for their ability to produce bioactive metabolites all strains were fermented and fermentation extracts profiled by LC-HRMS and tested for antimicrobial activity. Four strains exhibited antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus warneri. The production of secondary metabolites was assessed using a LC-HRMS-based metabolomics approach, where it was evaluated how differences in media composition can alter the production of chemical compounds. We used metabolomics as a tool to investigate the production of abyssomicins, an anticancer agent, and other secondary metabolites in three strains of the rare actinomycete Verrucosispora maris, all marine isolates from a sediment sample and associated to a starfish from the species Luidia senegalensis of Ubatuba – SP, Brazil. Nine different media compositions were evaluated and it was found that, among all strains, only RKMT_111 was capable of producing abyssomicins. The media composition study revealed that the production of abyssomicins was only achievable in BFM-11m. Although the three strains belong to the same species and the same location, it is worthwhile noticing that each isolate showed different capability for production of secondary metabolites. The products of fermentation of Erythrobacter vulgaris were evaluated using preparative HPLC, LC-HRMS and NMR techniques. Bacterial strain was isolated by drystamp method from a marine sediment sample from the coast of Ubatuba-SP, Brazil. After fully 16S rDNA sequence and identification, the marine isolate was fermented in large-scale, extracted and the compounds purified through HPLC-MS. Analysis of LC-HRMS and NMR of the isolated compounds led to the identification of two new cholic acid derivatives, 3- acetyl-glycocholic acid and 3-acetyl-glycodeoxycholic acid. Both new compounds may have been produced by the biotransformation of glycocholic acid and deoxycholic acid, respectively, already present in the cultivation medium. This is the first report of such compounds and also the first time an acylation has been observed for an Erythrobacter vulgaris marine isolate.
Descrição
Palavras-chave
Fauna marinha, Produtos naturais, Invertebrados marinhos, Bactérias, Actinobactéria, By-catch, Marine natural products, Invertebrates