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      Lignocellulose-Degrading Microbial Communities in Landfill Sites Represent a Repository of Unexplored Biomass-Degrading Diversity

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          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, 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.

          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.

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          Pretreatment technologies for an efficient bioethanol production process based on enzymatic hydrolysis: A review.

          Biofuel produced from lignocellulosic materials, so-called second generation bioethanol shows energetic, economic and environmental advantages in comparison to bioethanol from starch or sugar. However, physical and chemical barriers caused by the close association of the main components of lignocellulosic biomass, hinder the hydrolysis of cellulose and hemicellulose to fermentable sugars. The main goal of pretreatment is to increase the enzyme accessibility improving digestibility of cellulose. Each pretreatment has a specific effect on the cellulose, hemicellulose and lignin fraction thus, different pretreatment methods and conditions should be chosen according to the process configuration selected for the subsequent hydrolysis and fermentation steps. This paper reviews the most interesting technologies for ethanol production from lignocellulose and it points out several key properties that should be targeted for low-cost and advanced pretreatment processes. Copyright 2009 Elsevier Ltd. All rights reserved.
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            Environmental and Gut Bacteroidetes: The Food Connection

            Members of the diverse bacterial phylum Bacteroidetes have colonized virtually all types of habitats on Earth. They are among the major members of the microbiota of animals, especially in the gastrointestinal tract, can act as pathogens and are frequently found in soils, oceans and freshwater. In these contrasting ecological niches, Bacteroidetes are increasingly regarded as specialists for the degradation of high molecular weight organic matter, i.e., proteins and carbohydrates. This review presents the current knowledge on the role and mechanisms of polysaccharide degradation by Bacteroidetes in their respective habitats. The recent sequencing of Bacteroidetes genomes confirms the presence of numerous carbohydrate-active enzymes covering a large spectrum of substrates from plant, algal, and animal origin. Comparative genomics reveal specific Polysaccharide Utilization Loci shared between distantly related members of the phylum, either in environmental or gut-associated species. Moreover, Bacteroidetes genomes appear to be highly plastic and frequently reorganized through genetic rearrangements, gene duplications and lateral gene transfers (LGT), a feature that could have driven their adaptation to distinct ecological niches. Evidence is accumulating that the nature of the diet shapes the composition of the intestinal microbiota. We address the potential links between gut and environmental bacteria through food consumption. LGT can provide gut bacteria with original sets of utensils to degrade otherwise refractory substrates found in the diet. A more complete understanding of the genetic gateways between food-associated environmental species and intestinal microbial communities sheds new light on the origin and evolution of Bacteroidetes as animals’ symbionts. It also raises the question as to how the consumption of increasingly hygienic and processed food deprives our microbiota from useful environmental genes and possibly affects our health.
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              Biomass pretreatment: fundamentals toward application.

              Development of sustainable energy systems based on renewable biomass feedstocks is now a global effort. Lignocellulosic biomass contains polymers of cellulose, hemicellulose, and lignin, bound together in a complex structure. Liquid biofuels, such as ethanol, can be made from biomass via fermentation of sugars derived from the cellulose and hemicellulose within lignocellulosic materials, but the biomass must be subjected to pretreatment processes to liberate the sugars needed for fermentation. Production of value-added co-products along-side biofuels through integrated biorefinery processes creates the need for selectivity during pretreatment. This paper presents a survey of biomass pretreatment technologies with emphasis on concepts, mechanism of action and practicability. The advantages and disadvantages, and the potential for industrial applications of different pretreatment technologies are the highlights of this paper. Copyright © 2011 Elsevier Inc. All rights reserved.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                mSphere
                mSphere
                msph
                msph
                mSphere
                mSphere
                American Society for Microbiology (1752 N St., N.W., Washington, DC )
                2379-5042
                2 August 2017
                Jul-Aug 2017
                : 2
                : 4
                : e00300-17
                Affiliations
                [a ]School of Biological Sciences, Bangor University, Bangor, Gwynedd, United Kingdom
                [b ]Microbiology Research Group, Institute of Integrative Biology, University of Liverpool, Liverpool, United Kingdom
                [c ]Centre for Genomics Research, Institute of Integrative Biology, University of Liverpool, Liverpool, United Kingdom
                University of California, Berkeley
                Author notes
                Address correspondence to James E. McDonald, j.mcdonald@ 123456bangor.ac.uk .

                Citation Ransom-Jones E, McCarthy AJ, Haldenby S, Doonan J, McDonald JE. 2017. Lignocellulose-degrading microbial communities in landfill sites represent a repository of unexplored biomass-degrading diversity. mSphere 2:e00300-17. https://doi.org/10.1128/mSphere.00300-17.

                Author information
                http://orcid.org/0000-0002-6328-3752
                Article
                mSphere00300-17
                10.1128/mSphere.00300-17
                5541161
                28776044
                f0f684d9-8e9e-45e6-adad-20d6363f417e
                Copyright © 2017 Ransom-Jones et al.

                This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

                History
                : 7 July 2017
                : 19 July 2017
                Page count
                supplementary-material: 6, Figures: 3, Tables: 1, Equations: 0, References: 49, Pages: 13, Words: 8383
                Funding
                Funded by: RCUK | Biotechnology and Biological Sciences Research Council (BBSRC) https://doi.org/10.13039/501100000268
                Award ID: BB/L002043/1
                Award Recipient : James E. McDonald
                Categories
                Research Article
                Applied and Environmental Science
                Custom metadata
                July/August 2017

                bacteroidetes,biomass,cazymes,fibrobacter,firmicutes,landfill,cellulose degradation,cultivation,genomics,metagenomics,microbial ecology,spirochaetes

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