Potencial biotecnológico do metagenoma de rúmen bovino da raça nelore (Bos tauros indicus), visando à desconstrução da biomassa vegetal
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2017-06-22
Autores
Pavani, Claudio Damasceno [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A comunidade mundial busca pela obtenção de biocombustível celulósico, embora ainda seja um grande desafio a desconstrução do arranjo lignocelulósico para obtenção de açúcares livres de forma eficiente e economicamente viável. A fim de superar tais desafios, avanços conquistados pela metagenômica ressaltam sua aplicação como alternativa para compreender e desvendar o grande potencial metabólico presente nos ambientes, buscando encontrar em tais ambientes novas enzimas que atuem de forma eficiente na desconstrução do material lignocelulósico. Neste trabalho utilizou-se, a abordagem metagenômica para caracterização de genes com potencial para degradar biomassa vegetal e também para avaliação da diversidade taxonômica do ambiente ruminal bovino (Cow_1), bem como comparar os dados coletados com outros metagenomas disponíveis. Amostras de conteúdo ruminal de três bovinos machos da raça Nelore foram coletadas para a obtenção da amostra de DNA metagenômico, que posteriormente foi sequenciada pelo sequenciador HiScan SQ (Illumina). Foram obtidas aproximadamente 63 milhões de sequências (Cow _1), 2 x 100 pb. Dentre os 26 filos encontrados o filo Bacteroidetes foi o mais abundante, seguido de Firmicutes e Proteobacteria. Para a anotação gênica foi utilizado os bancos de dados Pfam, GO, KEGG e CAZy. Foram associadas ORFs relacionadas a 86 famílias de Glycoside Hydrolases (GHs), 52 famílias de Carbohydrate-Binding Modules (CBMs), 4 famílias de Auxiliary Activities (AAs), 16 famílias de Carbohydrate Esterases (CEs), 55 famílias de Glycosyl Transferases (GTs), 16 famílias de Polysaccharide Lyases (PLs), 1 família de S-layer homology (SLH) domain e 1 família de cohesina e dockerina, estes genes estão relacionados a formação do complexo enzimático para desconstrução da biomassa vegetal (celulossoma). O metagenoma da cow_1 analisado apresentou maior quantidade de genes relacionados à desconstrução da biomassa vegetal, quando comparados com outros 11 metagenomas. Além disso, os metagenomas dos ambientes lignocelulósicos apresentaram, principalmente, maior abundância de GH3, GH5, GH8, GH10 e GH11, genes que são importantes na degradação da lignocelulose quando comparados com o grupo de metagenomas não lignocelulósico. Os ambientes lignocelulósicos também revelaram maior abundância de sequências de domínios que se ligam na estrutura da celulose (CBM1, CBM10, CBM11, CBM16, CBM3, CBM63, CBM79 e CBM56), domínios que se ligam na estrutura da hemicelulose (CBM35, CBM61, CBM62 e CBM67) e domínios que atuam na estrutura da celulose e xilana (CBM37, CBM44 e CBM59). O metagenoma ruminal mostrou uma grande diversidade de enzimas responsáveis pela degradação da biomassa vegetal, demonstrando seu potencial como fonte de enzimas com potencial biotecnológico. Os metagenomas lignocelulósicos mostraram uma maior abundância de GH3, GH5, GH8, GH10 e GH11, genes que são importantes na degradação de celulose, celobiose, manana, xilana e xilo-oligossacarídeos quando comparados ao grupo não-lignocelulósico.
The world community search to obtain cellulosic biofuel; however the deconstruction of the lignocellulosic arrangement to obtain free sugars in an efficient and economically viable way is still a great challenge. Advances achieved through the metagenomic emphasize its application as an alternative to unfold the great metabolic potential present in the environments in order to overcome such challenges. Thus, a metagenomic approach was used to characterize genes with potential to degrade plant biomass and to evaluate the taxonomic diversity of the Nelore bovine ruminal environment. Samples were collected from three bovine males to obtain the DNA sample, which were sequentially sequenced by the HiScan SQ (Illumina) sequencer. About 63 million sequences, 2 x 100 bp, were obtained. Among the 26 phyla found, Bacteroidetes was the most abundant, followed by Firmicutes and Proteobacteria. For gene annotation was used the Pfam, GO, KEGG and CAZy databases. A total of 86 families of Glycoside Hydrolases (GHs), 52 families of Carbohydrate Binding Modules (CBMs), 4 families of Auxiliary Activities (AAs), 16 Esterase Carbohydrate (CEs), 55 GlycosylTransferases (GTs) Polysaccharide Lyases (PLs), 1 S-homology homology (SLH) domain and 1 family of cohesin and dockerine was associated. these genes are related to the formation of the enzymatic complex for deconstruction of plant biomass (cellulosome). The cow_1 metagenome presented a greater number of genes related to deconstruction of plant biomass when compared to other 11 metagenomes. In addition, the metagenomes of the lignocellulosic environments presented, mainly, greater abundance of GH3, GH5, GH8, GH10 and GH11, genes that are important in the degradation of lignocellulose when compared with the group of non-lignocellulosic metagenomes. Focusing on the CBMs, the metagenomes of the lignocellulosic group showed a greater abundance of CBM1, CBM10, CBM11, CBM16, CBM3, CBM63, CBM79 and CBM56 that bind in the cellulose structure; CBM35, CBM61, CBM62 and CBM67 which bind to the hemicellulose structure; CBM37, CBM44 and CBM59 that act on the structure of cellulose and xylan. The ruminal metagenome showed a great diversity of enzymes responsible for the degradation of the plant biomass, demonstrating its potential as a source of enzymes with biotechnological potential. Lignocellulosic metagenomes showed a greater abundance of GH3, GH5, GH8, GH10 and GH11, genes that are important in the degradation of cellulose, cellobiose, mannan, xylan and xylo-oligosaccharides when compared to the non-lignocellulosic group.
The world community search to obtain cellulosic biofuel; however the deconstruction of the lignocellulosic arrangement to obtain free sugars in an efficient and economically viable way is still a great challenge. Advances achieved through the metagenomic emphasize its application as an alternative to unfold the great metabolic potential present in the environments in order to overcome such challenges. Thus, a metagenomic approach was used to characterize genes with potential to degrade plant biomass and to evaluate the taxonomic diversity of the Nelore bovine ruminal environment. Samples were collected from three bovine males to obtain the DNA sample, which were sequentially sequenced by the HiScan SQ (Illumina) sequencer. About 63 million sequences, 2 x 100 bp, were obtained. Among the 26 phyla found, Bacteroidetes was the most abundant, followed by Firmicutes and Proteobacteria. For gene annotation was used the Pfam, GO, KEGG and CAZy databases. A total of 86 families of Glycoside Hydrolases (GHs), 52 families of Carbohydrate Binding Modules (CBMs), 4 families of Auxiliary Activities (AAs), 16 Esterase Carbohydrate (CEs), 55 GlycosylTransferases (GTs) Polysaccharide Lyases (PLs), 1 S-homology homology (SLH) domain and 1 family of cohesin and dockerine was associated. these genes are related to the formation of the enzymatic complex for deconstruction of plant biomass (cellulosome). The cow_1 metagenome presented a greater number of genes related to deconstruction of plant biomass when compared to other 11 metagenomes. In addition, the metagenomes of the lignocellulosic environments presented, mainly, greater abundance of GH3, GH5, GH8, GH10 and GH11, genes that are important in the degradation of lignocellulose when compared with the group of non-lignocellulosic metagenomes. Focusing on the CBMs, the metagenomes of the lignocellulosic group showed a greater abundance of CBM1, CBM10, CBM11, CBM16, CBM3, CBM63, CBM79 and CBM56 that bind in the cellulose structure; CBM35, CBM61, CBM62 and CBM67 which bind to the hemicellulose structure; CBM37, CBM44 and CBM59 that act on the structure of cellulose and xylan. The ruminal metagenome showed a great diversity of enzymes responsible for the degradation of the plant biomass, demonstrating its potential as a source of enzymes with biotechnological potential. Lignocellulosic metagenomes showed a greater abundance of GH3, GH5, GH8, GH10 and GH11, genes that are important in the degradation of cellulose, cellobiose, mannan, xylan and xylo-oligosaccharides when compared to the non-lignocellulosic group.
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Palavras-chave
Metagenoma, Degradação de lignocelulose, Rúmen, Biorefinaria, Biocombustível, Metagenome, Lignocellulose degradation, Rumen, Biorefinery, Biofuel