Identificação de novas enzimas de bibliotecas metagenômicas com aplicação biotecnológica para a agroindústria
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Data
2022-03-18
Autores
Lima, Natália Sarmanho Monteiro [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O desenvolvimento de novas tecnologias e novas abordagens para aumento da produtividade e melhoria das áreas cultivadas é uma busca constante. A utilização de enzimas de microrganismos é uma alternativa para alguns problemas encontrados no agronegócio, como utilização de resíduos agrícolas, controle biológico de pragas e doenças e biorremediação de defensivos agrícolas/pesticidas. As lacases e feruloil esterases são enzimas de grande importância biotecnológica. A metagenômica tem se mostrado bastante eficiente na busca de novos genes codificadores de proteínas/enzimas e a tecnologia do DNA recombinante facilita o acesso a esses genes. LacMeta é uma lacase metagenômica, que possui potencial para degradação de corantes, conforme resultados preliminares obtidos. Neste trabalho resolvemos aprofundar o estudo da aplicação biotecnológica desta enzima para biorremediação de outros compostos xenobióticos, visando determinar os parâmetros ótimos de reação bem como avaliar o potencial de detoxificação dos resíduos tratados. Para esta abordagem foram escolhidos o verde de malaquita (VM), amplamente utilizado na aquicultura apesar de proibições vigentes por diversas normas ambientais; o hidróxido de fentina (fungicida) e o imazapic (herbicida), ambos defensivos agrícolas largamente utilizados em monoculturas, visando verificar o potencial catalítico e mecanismos bioquímicos de atuação da LacMeta. Os resultados obtidos comprovaram a eficiência da LacMeta na detoxificação dos xenobióticos, sobretudo no que diz respeito a sua atuação na degradação completa do VM em 48 h, resultando em metabólitos inertes de acordo com os resultados de fitotoxidade e citotoxidade. Além disso, visando fomentar novos estudos de aplicação na degradação de resíduos agrícolas, prospectamos uma feruloil esterase (FAE) em bancos de dados metagenômicos. Esta enzima atua na degradação de ácidos hidroxinâmicos presentes na biomassa lignocelulósica. A nova FAE foi identificada como do tipo C. Análises preliminares da enzima em extrato celular solúvel bruto indicaram que a FeaC7 apresentou atividade enzimática positiva para esterase e tanase. Em conjunto, os resultados deste trabalho demonstram que, as enzimas metagenômicas LacMeta e FaeC7 possuem elevado potencial biotecnológico para aplicação na agroindústria, uma vez que podem atuar na degradação de resíduos agrícolas e na biorremediação de pesticidas.
The development of new technologies and new approaches to increase productivity and improve cultivated areas is a constant search. The use of enzymes from microorganisms is an alternative for some problems encountered in agribusiness, such as the use of agricultural residues, biological control of pests and diseases and bioremediation of agricultural pesticides/pesticides. Laccases and feruloyl esterases are enzymes of great biotechnological importance. Metagenomics has proved to be quite efficient in the search for new genes encoding proteins/enzymes, and recombinant DNA technology facilitates access to these genes. LacMeta is a metagenomic laccase, which has the potential for dye degradation, according to preliminary preliminary results obtained in my master's thesis. In this work we decided to deepen the study of the biotechnological application of this enzyme for the bioremediation of other xenobiotic compounds, aiming to determine the optimal reaction parameters as well as to evaluate the detoxification potential of the treated residues. Malachite green (VM) was chosen for this approach, widely used in aquaculture despite bans in force by various environmental authorities; fentin hydroxide (fungicide) and imazapic (herbicide), both pesticides widely used in monocultures with high persistence of environmental residues, aiming to verify the catalytic potential and biochemical mechanisms of action of LacMeta. The results obtained proved the efficiency of LacMeta in the detoxification of xenobiotics, especially with regard to its action in the complete degradation of VM in 48 h, resulting in inert metabolites according to the phytotoxicity and cytotoxicity results. Furthermore, in order to promote new application studies in the degradation of agricultural residues, we prospected a feruloyl esterase (FAE) in metagenomic databases, an enzyme that acts on the degradation of hydroxynamic acids present in lignocellulosic biomass. The new FAE was identified as type C. Preliminary analysis of the enzyme in crude soluble cell extract indicated that FeaC7 showed positive enzymatic activity for esterase and tannase. Together, the results of this work demonstrate that the metagenomic enzymes LacMeta and FaeC7 have a high biotechnological potential for application in the agroindustry, since they can act in the degradation of agricultural residues and in the bioremediation of pesticides.
The development of new technologies and new approaches to increase productivity and improve cultivated areas is a constant search. The use of enzymes from microorganisms is an alternative for some problems encountered in agribusiness, such as the use of agricultural residues, biological control of pests and diseases and bioremediation of agricultural pesticides/pesticides. Laccases and feruloyl esterases are enzymes of great biotechnological importance. Metagenomics has proved to be quite efficient in the search for new genes encoding proteins/enzymes, and recombinant DNA technology facilitates access to these genes. LacMeta is a metagenomic laccase, which has the potential for dye degradation, according to preliminary preliminary results obtained in my master's thesis. In this work we decided to deepen the study of the biotechnological application of this enzyme for the bioremediation of other xenobiotic compounds, aiming to determine the optimal reaction parameters as well as to evaluate the detoxification potential of the treated residues. Malachite green (VM) was chosen for this approach, widely used in aquaculture despite bans in force by various environmental authorities; fentin hydroxide (fungicide) and imazapic (herbicide), both pesticides widely used in monocultures with high persistence of environmental residues, aiming to verify the catalytic potential and biochemical mechanisms of action of LacMeta. The results obtained proved the efficiency of LacMeta in the detoxification of xenobiotics, especially with regard to its action in the complete degradation of VM in 48 h, resulting in inert metabolites according to the phytotoxicity and cytotoxicity results. Furthermore, in order to promote new application studies in the degradation of agricultural residues, we prospected a feruloyl esterase (FAE) in metagenomic databases, an enzyme that acts on the degradation of hydroxynamic acids present in lignocellulosic biomass. The new FAE was identified as type C. Preliminary analysis of the enzyme in crude soluble cell extract indicated that FeaC7 showed positive enzymatic activity for esterase and tannase. Together, the results of this work demonstrate that the metagenomic enzymes LacMeta and FaeC7 have a high biotechnological potential for application in the agroindustry, since they can act in the degradation of agricultural residues and in the bioremediation of pesticides.
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Palavras-chave
Lacase, Biorremediação, Microorganismos