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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Tomazetto, Geizecler [1, 2] ; Pimentel, Agnes C. [3] ; Wibberg, Daniel [4] ; Dixon, Neil [2] ; Squina, Fabio M. [1]
Total Authors: 5
[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
Total Affiliations: 4
Document type: Journal article
Source: Applied and Environmental Microbiology; v. 86, n. 18 SEP 2020.
Web of Science Citations: 1

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)

FAPESP's process: 18/23826-5 - Cellulosomes - a versatile enzymatic scaffold platform for bioconversion of lignocellulosic biomass into value-added fine chemicals
Grantee:Geizecler Tomazetto
Support type: Scholarships abroad - Research Internship - Post-doctor
FAPESP's process: 15/50590-4 - Lignin valorization in cellulosic ethanol plants: biocatalytic conversion via ferulic acid to high value chemicals
Grantee:Fábio Márcio Squina
Support type: Program for Research on Bioenergy (BIOEN) - Thematic Grants
FAPESP's process: 15/23279-6 - Metagenomic and metasecretomic analyses of an enrichment microbial consortium of anaerobic fungi to the degradation of sugarcane bagasse: prospection of cellulosomas and lignocellolytic enzymes
Grantee:Geizecler Tomazetto
Support type: Scholarships in Brazil - Post-Doctorate
FAPESP's process: 16/01926-2 - Influence of an accessory domain in biochemical and structural characteristics of the cellulase CelE2
Grantee:Agnes Cristina Pimentel
Support type: Scholarships in Brazil - Master