Otimização do cultivo da actinobactéria marinha Micromonospora sp. BRB-607 por planejamento fatorial para obtenção de extrato com potencial anticâncer
Data
2023-07-07
Autores
Sábio, João Vitor 
Silva, João Vitor Sábio da
Orientadores
Jimenez, Paula Christine
Tipo
Trabalho de conclusão de curso
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Doenças como o câncer ainda não possuem tratamentos completamente efetivos e seletivos, o que torna necessária a contínua bioprospecção de fármacos isolados de bactérias. Estes micro-organismos são uma fonte promissora de princípios ativos com potencial farmacológico, sendo que o gênero Micromonospora, particularmente, abrange actinobactérias de onde já foram obtidos diversos fármacos que hoje encontram-se disponíveis no mercado. O suprimento destas moléculas obtidas de microrganismos é um dos maiores desafios do desenvolvimento de fármacos, assim é importante que se busque formas de otimizar o processo de obtenção destes produtos a partir dos micro-organismos produtores. Planejamento fatorial é um método estatístico usado para estudar o efeito de múltiplas variáveis em uma resposta específica, permitindo a identificação de condições de cultura que melhoram a produção de metabólitos. Esta é uma ferramenta amplamente utilizada para otimizar processos bacterianos visando metabólitos primários, porém não é comumente aplicado quando os alvos são metabólitos secundários. Aqui, aplicamos o planejamento fatorial para investigar parâmetros de cultura que afetam a produção de compostos citotóxicos pela cepa de actinobactérias Micromonospora sp. BRB-607, recuperado de sedimentos marinhos de Ubatuba, São Paulo, Brasil. Um protocolo de planejamento fatorial fracionário 2 5-1 foi aplicado para avaliar as condições de cultura que afetam o potencial citotóxico dos extratos de BRB-607. O crescimento da biomassa e o rendimento do extrato variaram entre 1,9 a 6,7 g/L e 1,6 a 37,2% entre as diferentes condições, enquanto os pontos centrais foram em média de 5,5 g/L e 7%, respectivamente. A citotoxicidade variou entre 1,24 e > 50μg/mL. Amido e pH, salinidade e peptona foram significativos e diretamente proporcionais ao rendimento de biomassa e extrato, respectivamente. O extrato de levedura foi significativo e inversamente proporcional ao rendimento do extrato e à citotoxicidade. Além disso, a citotoxicidade dos extratos também foi significativamente afetada pela salinidade (inversamente) e pH (diretamente). Esses resultados revelam que o planejamento fatorial é uma ferramenta viável para entender melhor os parâmetros que afetam a produção de metabólitos secundários microbianos e, além disso, que melhores condições de cultivo são importantes para permitir o isolamento e posterior caracterização da química e da atividade biológica dos princípios ativos obtidos de Micromonospora sp. BRB-607.
Diseases such as cancer still do not have completely effective and selective treatments, which makes the continuous bioprospecting of drugs isolated from bacteria necessary. These microorganisms are a promising source of active principles with pharmacological potential, and the genus Micromonospora, in particular, covers actinobacteria from which several drugs that are currently available on the market have already been obtained. The supply of these molecules obtained from microorganisms is one of the biggest challenges in drug development, so it is important to seek ways to optimize the process of obtaining these products from the producing microorganisms. Factorial design is a statistical method used to study the effect of multiple variables on a specific response, allowing for identification of culture conditions that improve metabolite production. Factorial design is a widely used tool for optimizing bacterial processes targeting primary metabolites, however it is not commonly applied when the targets are secondary metabolites. Herein, we applied factorial design to investigate culture parameters that affect the production of cytotoxic compounds by actinobacteria strain Micromonospora sp. BRB-607, recovered from marine sediments from Ubatuba, São Paulo, Brazil. A 2 5-1 fractional factorial design protocol was applied to evaluate culture conditions affecting the cytotoxic potential of BRB607 extracts. Biomass growth and extract yield ranged between 1.9 to 6.7 g/L and 1.6 to 37.2% among the different conditions, while center points averaged 5.5 g/L and 7%, respectively. Cytotoxicity varied between 1.24 and > 50μg/mL. Starch and pH, and salinity and peptone, were significant and directly proportional to biomass and extract yield, respectively. Yeast extract was significant and inversely proportional to both extract yield and cytotoxicity. Furthermore, cytotoxicity of extracts was also significantly affected by salinity (inversely) and pH (directly). These results reveal factorial design as a viable tool to better understand the parameters affecting production of microbial secondary metabolites and, moreover, that improved culture conditions are important to enable isolation and further characterization of the chemistry and biological activity of the active principles obtained from Micromonospora sp. BRB-607.
Diseases such as cancer still do not have completely effective and selective treatments, which makes the continuous bioprospecting of drugs isolated from bacteria necessary. These microorganisms are a promising source of active principles with pharmacological potential, and the genus Micromonospora, in particular, covers actinobacteria from which several drugs that are currently available on the market have already been obtained. The supply of these molecules obtained from microorganisms is one of the biggest challenges in drug development, so it is important to seek ways to optimize the process of obtaining these products from the producing microorganisms. Factorial design is a statistical method used to study the effect of multiple variables on a specific response, allowing for identification of culture conditions that improve metabolite production. Factorial design is a widely used tool for optimizing bacterial processes targeting primary metabolites, however it is not commonly applied when the targets are secondary metabolites. Herein, we applied factorial design to investigate culture parameters that affect the production of cytotoxic compounds by actinobacteria strain Micromonospora sp. BRB-607, recovered from marine sediments from Ubatuba, São Paulo, Brazil. A 2 5-1 fractional factorial design protocol was applied to evaluate culture conditions affecting the cytotoxic potential of BRB607 extracts. Biomass growth and extract yield ranged between 1.9 to 6.7 g/L and 1.6 to 37.2% among the different conditions, while center points averaged 5.5 g/L and 7%, respectively. Cytotoxicity varied between 1.24 and > 50μg/mL. Starch and pH, and salinity and peptone, were significant and directly proportional to biomass and extract yield, respectively. Yeast extract was significant and inversely proportional to both extract yield and cytotoxicity. Furthermore, cytotoxicity of extracts was also significantly affected by salinity (inversely) and pH (directly). These results reveal factorial design as a viable tool to better understand the parameters affecting production of microbial secondary metabolites and, moreover, that improved culture conditions are important to enable isolation and further characterization of the chemistry and biological activity of the active principles obtained from Micromonospora sp. BRB-607.
Descrição
Citação
SÁBIO, João Vitor. Otimização do cultivo da actinobactéria marinha Micromonospora sp. BRB-607 por planejamento fatorial para obtenção de extrato com potencial anticâncer. 2023. 31 f. Trabalho de conclusão de curso (Graduação em Bacharelado Interdisciplinar em Ciência e Tecnologia do Mar) - Instituto do Mar, Universidade Federal de São Paulo, Santos, 2023.