Explorando uma comunidade bacteriana isolada de uma pilha de bagaço de cana-de-açúcar e seu potencial funcional na degradação de biomassa lignocelulósica
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Data
2018-02-26
Autores
Funnicelli, Michelli Inácio Gonçalves
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Universidade Estadual Paulista (Unesp)
Resumo
RESUMO- Os biocombustíveis são uma alternativa atraente para a substituição de combustíveis fósseis que impulsionaram o interesse global na conversão de biomassa lignocelulósica. Diversos trabalhos são realizados para otimização das etapas para produção de etanol de segunda geração em termos de custo global para o processo. A demanda pela busca de enzimas específicas utilizadas em processos industriais tem impulsionado a prospecção de novas enzimas microbianas com capacidade de desconstruir biomassa vegetal. Dentre elas, as enzimas do grupo das carboidrases são as principais atuantes envolvidas no processo de conversão de biomassa. Diante disso, foi realizada a caracterização de uma comunidade microbiana denominada CB10b, que tem potencialidade em degradar a biomassa. Os estudos foram realizados a partir do sequenciamento total do DNA obtido da comunidade e da prospecção de genes úteis nos processos de conversão de biomassa vegetal. Foram isoladas 6 bactérias pertencentes aos respectivos gêneros: Chitinophaga (que apresentou atividade enzimática em carboximetilcelulose), Pandoraea e Labrys, identificado pelas análises do gene 16S rRNA e da região espaçadora intergênica (ITS) entre os genes 16S-23S rRNA. Por meio das análises dos genes foi possível identificar 16.340 ORFs, sendo anotadas 624 ORFs associadas ao banco de dados do CAZy, distribuídas entre as classes glicosil hidrolases (GHs) com 37,98% (237 ORFs); glicosil transferases (GTs) com 21,63% (135 ORFs); polissacarídeo liases (PLs) com 3,18% (20 ORFs); carboidrato esterases (CEs) com 17,00% (106 ORFs); atividades auxiliares (AAs) com 4,81% (30 ORFs) e módulos de ligação a carboidratos (CBMs) com 15,38% (96 ORFs). Adicionalmente foi possível recuperar através da análise do metagenoma, o genoma parcial de uma bactéria do gênero Pandoraea. Deste modo, o estudo com a comunidade bacteriana CB10b apresentou genes envolvidos a degradação de biomassa lignocelulósica.
ABSTRACT- Biofuels are an attractive alternative for replacing fossil fuels, bringing the world`s attention to the conversion processes of lignocellulosic biomass. Multiple works are currently being done with to optimize the steps in second-generation ethanol production, seeking to reduce the global costs in this process. Demand for specific microbial enzymes is driving the prospection of novel ones able to degrade plant biomass. Among those, carbohydrases are the protagonists in these processes. In this context, we characterized a microbial community called CB10b and tested it`s potential to degrade biomass. We performed studies on this community by total DNA sequencing followed by prospecting genes related to conversion of plant biomass. Six bacteria were isolated, belonging to the genera: Chitinophaga, who presented enzymatic activity on for carboxymethyl cellulose; Pandoraea and Labrys, identified through 16S rRNA and 16S-23S rRNA internal transcribed spacer (ITS) gene analysis. Some 16,340 ORFs were identified, of which 624 were associated with CAZy database, 37.98% (237 ORFs) classified as glycosyl hydrolases (GHs); 21.63% (135 ORFs) as glycosyltransferases (GTs); 17.00% (106 ORFs) were carbohydrate esterases (CEs); 15.38% (96 ORFs) as carbohydrate-binding modules; 4.81% (30 ORFs) as auxiliary activities (AAs) and 3.18% (20 ORFs) as polysaccharide lyase. In addition, it was possible to recover by metagenome analysis the partial genome of a bacterium of the genus Pandoraea. Thus, the study with the bacterial community CB10b presented genes involved the degradation of lignocellulosic biomass.
ABSTRACT- Biofuels are an attractive alternative for replacing fossil fuels, bringing the world`s attention to the conversion processes of lignocellulosic biomass. Multiple works are currently being done with to optimize the steps in second-generation ethanol production, seeking to reduce the global costs in this process. Demand for specific microbial enzymes is driving the prospection of novel ones able to degrade plant biomass. Among those, carbohydrases are the protagonists in these processes. In this context, we characterized a microbial community called CB10b and tested it`s potential to degrade biomass. We performed studies on this community by total DNA sequencing followed by prospecting genes related to conversion of plant biomass. Six bacteria were isolated, belonging to the genera: Chitinophaga, who presented enzymatic activity on for carboxymethyl cellulose; Pandoraea and Labrys, identified through 16S rRNA and 16S-23S rRNA internal transcribed spacer (ITS) gene analysis. Some 16,340 ORFs were identified, of which 624 were associated with CAZy database, 37.98% (237 ORFs) classified as glycosyl hydrolases (GHs); 21.63% (135 ORFs) as glycosyltransferases (GTs); 17.00% (106 ORFs) were carbohydrate esterases (CEs); 15.38% (96 ORFs) as carbohydrate-binding modules; 4.81% (30 ORFs) as auxiliary activities (AAs) and 3.18% (20 ORFs) as polysaccharide lyase. In addition, it was possible to recover by metagenome analysis the partial genome of a bacterium of the genus Pandoraea. Thus, the study with the bacterial community CB10b presented genes involved the degradation of lignocellulosic biomass.
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Palavras-chave
Biocombustíveis, Biomassa, Chitinophaga, Labrys, Pandoraea, biofuels, biomass