Multi-omic Directed Discovery of Cellulosomes, Polysaccharide Utilization Loci, and Lignocellulases from an Enriched Rumen Anaerobic Consortium

Autor(es)

Tomazetto, Geizecler ^{[1, 2]} ; Pimentel, Agnes C. ^[3] ; Wibberg, Daniel ^[4] ; Dixon, Neil ^[2] ; Squina, Fabio M. ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sorocaba, Programa Proc Tecnol & Ambientais, Sorocaba - Brazil

^[2] Univ Manchester, Manchester Inst Biotechnol, Dept Chem, Manchester, Lancs - England

^[3] Univ Estadual Campinas, UNICAMP, Inst Biol IB, Dept Bioquim, Campinas, SP - Brazil

^[4] Bielefeld Univ, Ctr Biotechnol CeBiTec, Genome Res Ind Microorganisms, Bielefeld - Germany

• Tipo de documento: Artigo Científico
• Fonte: Applied and Environmental Microbiology; v. 86, n. 18 SEP 2020.
• Citações Web of Science: 1

Resumo

Lignocellulose is one of the most abundant renewable carbon sources, representing an alternative to petroleum for the production of fuel and chemicals. Nonetheless, the lignocellulose saccharification process, to release sugars for downstream applications, is one of the most crucial factors economically challenging to its use. The synergism required among the various carbohydrate-active enzymes (CAZymes) for efficient lignocellulose breakdown is often not satisfactorily achieved with an enzyme mixture from a single strain. To overcome this challenge, enrichment strategies can be applied to develop microbial communities with an efficient CAZyme arsenal, incorporating complementary and synergistic properties, to improve lignocellulose deconstruction. We report a comprehensive and deep analysis of an enriched rumen anaerobic consortium (ERAC) established on sugarcane bagasse (SB). The lignocellulolytic abilities of the ERAC were confirmed by analyzing the depolymerization of bagasse by scanning electron microscopy, enzymatic assays, and mass spectrometry. Taxonomic analysis based on 16S rRNA sequencing elucidated the community enrichment process, which was marked by a higher abundance of Firmicutes and Synergistetes species. Shotgun metagenomic sequencing of the ERAC disclosed 41 metagenome-assembled genomes (MAGs) harboring cellulosomes and polysaccharide utilization loci (PULs), along with a high diversity of CAZymes. The amino acid sequences of the majority of the predicted CAZymes (60% of the total) shared less than 90% identity with the sequences found in public databases. Additionally, a clostridial MAG identified in this study produced proteins during consortium development with scaffoldin domains and CAZymes appended to dockerin modules, thus representing a novel cellulosome-producing microorganism. IMPORTANCE The lignocellulolytic ERAC displays a unique set of plant polysaccharide-degrading enzymes (with multimodular characteristics), cellulosomal complexes, and PULs. The MAGs described here represent an expansion of the genetic content of rumen bacterial genomes dedicated to plant polysaccharide degradation, therefore providing a valuable resource for the development of biocatalytic toolbox strategies to be applied to lignocellulose-based biorefineries. (AU)

• Processo FAPESP: 16/01926-2 - Influência de um domínio acessório nas características bioquímicas e estruturais da celulase CelE2
• Beneficiário: Agnes Cristina Pimentel
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 18/23826-5 - Celulossomas: uma versátil plataforma enzimática para a bioconversão de biomassa lignocelulósica em produtos químicos de valor agregado
• Beneficiário: Geizecler Tomazetto
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado
• Processo FAPESP: 15/50590-4 - Valorização da lignina em plantas de etanol celulósico: a conversão biocatalítica via ácido ferúlico a produtos químicos de alto valor
• Beneficiário: Fábio Márcio Squina
• Modalidade de apoio: Auxílio à Pesquisa - Programa BIOEN - Temático
• Processo FAPESP: 15/23279-6 - Análise metagenômica e metasecretômica de um consórcio microbiano enriquecido de fungos anaeróbicos degradador de bagaço de cana: prospecção de celulossomas e enzimas lignocelulolíticas
• Beneficiário: Geizecler Tomazetto
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado