Abstract

The microbial conversion of lignocellulosic biomass for biofuel production represents a renewable alternative to fossil fuels. However, the discovery of new microbial enzymes with high activity is critical for improving biomass conversion processes. While attempts to identify superior lignocellulose-degrading enzymes have focused predominantly on the animal gut, biomass-degrading communities in landfill sites represent an unexplored resource of hydrolytic enzymes for biomass conversion. Here, to address the paucity of information on biomass-degrading microbial diversity beyond the gastrointestinal tract, cellulose (cotton) "baits" were incubated in landfill leachate microcosms to enrich the landfill cellulolytic microbial community for taxonomic and functional characterization. Metagenome and 16S rRNA gene amplicon sequencing demonstrated the dominance of Firmicutes, Bacteroidetes, Spirochaetes, and Fibrobacteres in the landfill cellulolytic community. Functional metagenome analysis revealed 8,371 carbohydrate active enzymes (CAZymes) belonging to 244 CAZyme families. In addition to observing biomass-degrading enzymes of anaerobic bacterial "cellulosome" systems of members of the Firmicutes, we report the first detection of the Fibrobacter cellulase system and the Bacteroidetes polysaccharide utilization locus (PUL) in landfill sites. These data provide evidence for the presence of multiple mechanisms of biomass degradation in the landfill microbiome and highlight the extraordinary functional diversity of landfill microorganisms as a rich source of biomass-degrading enzymes of potential biotechnological significance. IMPORTANCE The microbial conversion of lignocellulosic biomass for biofuel production represents a renewable alternative to fossil fuels. However, the discovery of new microbial enzymes with high activity is critical for improving biomass conversion processes. While attempts to identify superior lignocellulose-degrading enzymes have focused predominantly on the animal gut, biomass-degrading communities in landfill sites represent an unexplored resource of hydrolytic enzymes for biomass conversion. Here, we identified Firmicutes, Spirochaetes, and Fibrobacteres as key phyla in the landfill cellulolytic community, detecting 8,371 carbohydrate active enzymes (CAZymes) that represent at least three of the recognized strategies for cellulose decomposition. These data highlight substantial hydrolytic enzyme diversity in landfill sites as a source of new enzymes for biomass conversion.

Highlights

  • The microbial conversion of lignocellulosic biomass for biofuel production represents a renewable alternative to fossil fuels

  • The aims of our study were to (i) utilize 16S rRNA gene amplicon sequencing of replicated raw leachate and cellulose enrichment microcosm samples to identify members of the landfill microbiome that are significantly enriched with cellulose amendment and (ii) to identify the functional diversity and taxonomic identity of the landfill biomass-degrading microbiome using deep metagenomic sequencing and taxonomic binning

  • The community composition of DNA extracted from three raw leachate samples and nine replicate landfill leachate microcosms containing 1% Avicel was determined using 16S rRNA gene amplicon sequencing on the Ion Torrent PGM platform

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Summary

Introduction

The microbial conversion of lignocellulosic biomass for biofuel production represents a renewable alternative to fossil fuels. We identified Firmicutes, Spirochaetes, and Fibrobacteres as key phyla in the landfill cellulolytic community, detecting 8,371 carbohydrate active enzymes (CAZymes) that represent at least three of the recognized strategies for cellulose decomposition. These data highlight substantial hydrolytic enzyme diversity in landfill sites as a source of new enzymes for biomass conversion. Recent attempts to identify superior lignocellulose-degrading enzymes and microorganisms have focused predominantly on anaerobic gut environments such as the bovine rumen (4), elephant gut (5), and termite gut (6), in addition to biogas reactors (7) These environments harbor microbial communities that have evolved to attack lignocellulosic biomass without the pretreatments currently used in commercial processes (8). Members of the Chlamydiae/Verrucomicrobia group (17, 18), the Cytophaga–Flexibacter–Bacteroides group (17, 18), and phyla Planctomycetes (17), TM6 (19), Chloroflexi (19, 20), Actinobacteria (19), Proteobacteria (19, 20), Lentisphaerae (20), Spirochaetes (20), Synergistetes (20), Thermotogae (20), and Fibrobacteres (21) are detected

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