Otimização do cultivo da bactéria marinha actinomadura sp. BRA-177 por planejamento fatorial para obtenção de prodigininas
Data
2023-12-08
Autores
Matsuda, Vitória Ami 
Orientadores
Jimenez, Paula Christine
Tipo
Trabalho de conclusão de curso
Título da Revista
ISSN da Revista
Título de Volume
Resumo
Os desafios que microorganismos marinhos enfrentam nos oceanos estimularam o desenvolvimento de estratégias metabólicas singulares e de alta capacidade adaptativa. Particularmente as actinobactérias marinhas, são potenciais fontes de metabólitos secundários de interesse para as empresas e indústrias farmacêuticas para o desenvolvimento de fármacos produzidos a partir de seus produtos naturais. O desenvolvimento de novos fármacos é um processo caro (em torno de 1,3 bilhão e 1,8 bilhão de dólares por medicamento), demorado (média de 10 a 15 anos) e envolve várias etapas, desde a pesquisa pré-clínica até seu registro no órgão de fiscalização sanitária. A bioprospecção é uma investigação sistemática da biodiversidade na busca de recursos com valor econômico e social. Trabalhos anteriores identificaram um extrato bioativo obtido da cepa de bactéria marinha Actinomadura sp. BRA-177, recuperada de sedimento coletado em expedição ao Arquipélago de São Pedro e São Paulo e armazenada no banco de cepas do grupo de pesquisa. Esta cepa é produtora de prodigininas, moléculas com alto potencial anticâncer e de grande interesse do grupo de pesquisa. Entretanto, a disponibilidade de maiores quantidades deste extrato com atividade biológica e da moléculas para a continuação da pesquisa e desenvolvimento dos fármacos de origem natural ainda é um obstáculo a ser superado. Portanto, o objetivo deste trabalho foi otimizar a obtenção de extratos biologicamente ativos, alterando as condições do meio de cultivo da cepa BRA-177 através de um protocolo de planejamento fatorial. Visando compreender as condições ideais para o cultivo da respectiva cepa de modo a obter os metabólitos bioativos com mais eficiência, foi aplicado o método do planejamento fatorial fracionado 25-1, onde foram consideradas como variáveis resposta biomassa, rendimento e citotoxicidade do extrato. Para a biomassa e IC50, a variável significativa foi o pH, enquanto para o rendimento de extrato foi a salinidade. Quantitativamente, a biomassa variou de 0,0473 g/mL a 0,2038 g/mL, o rendimento de extrato foi entre 13,4% e 40,4%, e IC50 foi de 0,41 a 52,01 μg/mL. Este estudo mostrou que o planejamento fatorial é uma ferramenta útil para entender como os parâmetros de cultivo influenciam na produção de extratos bioativos.
The challenges that marine microorganisms face in the oceans have stimulated the development of unique metabolic strategies and high adaptive capacity. Marine actinobacteria, in particular, are potential sources of secondary metabolites of interest to pharmaceutical companies and industries for the development of drugs produced from their natural products. The development of new drugs is an expensive process (around 1.3 billion and 1.8 billion dollars per drug), time consuming (average of 10 to 15 years) and involves several steps, from pre-clinical research to their registration with the health surveillance agency. Bioprospecting is a systematic investigation of biodiversity in the search for resources with economic and social value.Previous work has identified a bioactive extract obtained from the marine bacterium strain Actinomadura sp. BRA-177, recovered from sediment collected during an expedition to the Archipelago of São Pedro and São Paulo and stored in the research group's strain bank. This strain is a producer of prodigynins, molecules with high anticancer potential and of great interest to the research group. However, the availability of larger quantities of this extract with biological activity and of the molecules for further research and development of drugs of natural origin is still an obstacle to be overcome. Therefore, the objective of this work was to optimize the obtaining of biologically active extracts by changing the conditions of the BRA-177 strain culture medium through a factorial design protocol. Aiming to understand the ideal conditions for cultivating the respective strain in order to obtain bioactive metabolites more efficiently, the 25-1 fractional factorial planning method was applied, where biomass, yield and cytotoxicity of the extract were considered as response variables. For biomass and IC50, the significant variable was pH, while for extract yield it was salinity. Quantitatively, biomass ranged from 0.0473 g/mL to 0.2038 g/mL, extract yield was between 13.4% and 40.4%, and IC50 was from 0.41 to 52.01 μg/mL. This study showed that factorial design is a useful tool for understanding how cultivation parameters influence the production of bioactive extracts.
The challenges that marine microorganisms face in the oceans have stimulated the development of unique metabolic strategies and high adaptive capacity. Marine actinobacteria, in particular, are potential sources of secondary metabolites of interest to pharmaceutical companies and industries for the development of drugs produced from their natural products. The development of new drugs is an expensive process (around 1.3 billion and 1.8 billion dollars per drug), time consuming (average of 10 to 15 years) and involves several steps, from pre-clinical research to their registration with the health surveillance agency. Bioprospecting is a systematic investigation of biodiversity in the search for resources with economic and social value.Previous work has identified a bioactive extract obtained from the marine bacterium strain Actinomadura sp. BRA-177, recovered from sediment collected during an expedition to the Archipelago of São Pedro and São Paulo and stored in the research group's strain bank. This strain is a producer of prodigynins, molecules with high anticancer potential and of great interest to the research group. However, the availability of larger quantities of this extract with biological activity and of the molecules for further research and development of drugs of natural origin is still an obstacle to be overcome. Therefore, the objective of this work was to optimize the obtaining of biologically active extracts by changing the conditions of the BRA-177 strain culture medium through a factorial design protocol. Aiming to understand the ideal conditions for cultivating the respective strain in order to obtain bioactive metabolites more efficiently, the 25-1 fractional factorial planning method was applied, where biomass, yield and cytotoxicity of the extract were considered as response variables. For biomass and IC50, the significant variable was pH, while for extract yield it was salinity. Quantitatively, biomass ranged from 0.0473 g/mL to 0.2038 g/mL, extract yield was between 13.4% and 40.4%, and IC50 was from 0.41 to 52.01 μg/mL. This study showed that factorial design is a useful tool for understanding how cultivation parameters influence the production of bioactive extracts.
Descrição
Citação
MATSUDA, Vitória Ami. Otimização do Cultivo da Bactéria Marinha Actinomadura sp. BRA-177 por Planejamento Fatorial para Obtenção de Prodigininas. 2023. 33 f. Trabalho de conclusão de curso (Graduação em Engenharia Ambiental) - Universidade Federal de São Paulo, Instituto do Mar, Santos, 2023.