Cellulose Deconstruction Research Articles

Heterologously Expressed Cellobiose Dehydrogenase Acts as Efficient Electron-Donor of Lytic Polysaccharide Monooxygenase for Cellulose Degradation in Trichoderma reesei.

The conversion of lignocellulosic biomass to second-generation biofuels through enzymes is achieved at a high cost. Filamentous fungi through a combination of oxidative enzymes can easily disintegrate the glycosidic bonds of cellulose. The combination of cellobiose dehydrogenase (CDH) with lytic polysaccharide monooxygenases (LPMOs) enhances cellulose degradation in many folds. CDH increases cellulose deconstruction via coupling the oxidation of cellobiose to the reductive activation of LPMOs by catalyzing the addition of oxygen to C-H bonds of the glycosidic linkages. Fungal LPMOs show different regio-selectivity (C1 or C4) and result in oxidized products through modifications at reducing as well as nonreducing ends of the respective glucan chain. T. reesei LPMOs have shown great potential for oxidative cleavage of cellobiose at C1 and C4 glucan bonds, therefore, the incorporation of heterologous CDH further increases its potential for biofuel production for industrial purposes at a reduced cost. We introduced CDH of Phanerochaete chrysosporium (PcCDH) in Trichoderma reesei (which originally lacked CDH). We purified CDH through affinity chromatography and analyzed its enzymatic activity, electron-donating ability to LPMO, and the synergistic effect of LPMO and CDH on cellulose deconstruction. The optimum temperature of the recombinant PcCDH was found to be 45 °C and the optimum pH of PcCDH was observed as 4.5. PcCDH has high cello-oligosaccharide kcat, Km, and kcat/Km values. The synergistic effect of LPMO and cellulase significantly improved the degradation efficiency of phosphoric acid swollen cellulose (PASC) when CDH was used as the electron donor. We also found that LPMO undergoes auto-oxidative inactivation, and when PcCDH is used an electron donor has the function of a C1-type LPMO electron donor without additional substrate increments. This work provides novel insights into finding stable electron donors for LPMOs and paves the way forward in discovering efficient CDHs for enhanced cellulose degradation.

Aspergillus fumigatus Lytic Polysaccharide Monooxygenase AfLPMO9D: Biochemical Properties and Photoactivation of a Multi-Domain AA9 Enzyme

Lytic polysaccharide monooxygenases (LPMOs) are critical players in enzymatic deconstruction of cellulose. A number of LPMOs have been identified at a genomics level; however, they still need to be characterized and validated for use in industrial processes aimed at cellulose deconstruction. In the present study, we biochemically characterized a new LPMO, a member of auxiliary activities family 9 (AA9) from the filamentous fungus Aspergillus fumigatus (AfLPMO9D). This LPMO demonstrated higher efficiency against amorphous cellulose as compared to more recalcitrant forms of cellulose such as bacterial cellulose and Avicel. AfLPMO9D has a capacity to oxidize the substrate at either the C1 or C4 positions, with pH-dependent regioselectivity. Photoactivation experiments demonstrated that light-stimulated chlorophyllin triggers AfLPMO9D activation without requirements of an external electron donor. AfLPMO9D is capable of boosting phosphoric acid-swollen cellulose depolymerization via GH7 endoglucanase and cellobiohydrolase. The results of the present study might help to elucidate the role of different LPMOs in cellulosic fiber deconstruction.

Systematic bioprospection for cellulolytic actinomycetes in the Chihuahuan Desert: isolation and enzymatic profiling.

The quest for microbial cellulases has intensified as a response to global challenges in biofuel production. The efficient deconstruction of lignocellulosic biomass holds promise for generating valuable products in various industries such as food, textile, and detergents. This article presents a systematic bioprospection aimed at isolating actinomycetes with exceptional cellulose deconstruction capabilities. Our methodology explored the biodiverse oligotrophic region of Cuatro Cienegas, Coahuila, within the Chihuahuan Desert. Among the evaluated actinomycetes collection, 78% exhibited cellulolytic activity. Through a meticulous screening process based on enzymatic index evaluation, we identified a highly cellulolytic Streptomyces strain for further investigation. Submerged fermentation of this strain revealed an endoglucanase enzymatic activity of 149 U/mg. Genomic analysis of strain Streptomyces sp. STCH565-A revealed unique configurations of carbohydrate-active enzyme (CAZyme) genes, underscoring its potential for lignocellulosic bioconversion applications. These findings not only highlight the significance of the Chihuahuan Desert as a rich source of cellulolytic microorganisms but also offer insights into the systematic exploration and selection of high-performing cellulolytic microorganisms for application in diverse environmental contexts. In conclusion, our bioprospecting study lays a foundation for harnessing the cellulolytic potential of actinomycetes from the Chihuahuan Desert, with implications for advancing cellulose deconstruction processes in various industries. The findings can serve as a blueprint for future bioprospecting efforts in different regions, facilitating the targeted discovery of microorganisms with exceptional cellulosic deconstruction capabilities.

Elucidating multi-scale deconstruction of bamboo crystalline cellulose by novel acetylcholine chloride based deep eutectic solvents for enhanced enzymatic hydrolysis

Exploration and application of novel and sustainable deep eutectic solvents (DESs) on lignocellulose pretreatment are promising approaches to efficient bioconversion. However, comprehensive understanding on efficient DESs selection and inner mechanisms of lignocellulose deconstruction are still limited. Herein, bio-based acetylcholine chloride (AChCl) was firstly utilized as hydrogen bond acceptor in cooperation with lactic acid, urea and glycerol respectively for microwave-assisted pretreatment on moso bamboo in short time. High enzymatic conversion yield of bamboo cellulose (∼85%) was achieved by lactic acid-AChCl DES due to its lower pH value and viscosity, followed by glycerol-AChCl and urea-AChCl DES. In addition to the features of proposed DESs, changed chemical and physical properties of highly recalcitrant bamboo biomass after pretreatments were systematically determined. Besides of great amount of lignin (∼90%)/ xylan (100%) removal, micro- to nano-scale variations of cellulose structures including more dissociated microfibrils, lowered crystallinity, and enlarged crystallite sizes all played key roles in promoting bioconversion yields. Moreover, the multi-factors contributing to improved hydrolysis performance were integrated to evaluate the effects of novel DESs pretreatments on lignocellulose deconstruction, which may provide new strategies on bioconversion enhancement and process optimization.

Insights into cellulose deconstruction and pseudo-lignin formation during deep eutectic solvent treatment

Insights into cellulose deconstruction and pseudo-lignin formation during deep eutectic solvent treatment

Catalytic Wood Fractionation into Chemicals in Supercritical Ethanol and n-Heptane: Potential and Limitations

Direct selective wood fractionation into chemicals is an approach that has attracted recent attention. The application of sub- and supercritical (SC) alcohols to fractionate wood into solid cellulose and liquefy phenolic monomers is a process now widely known as “lignin first”. It is justified to study the potential of other SC organic solvents of variable polarities. Herein, we compare the abilities of SC ethanol and SC n-heptane to fractionate pine wood near their critical point. While near-critical ethanol has more affinity for lignin fraction, we show that near-critical n-heptane has preference for carbohydrate deconstruction. If SC ethanol favors biooil formation which contains important ethyl/ethoxy groups, the alkane greatly favors solid carbon products. The impact of addition of heterogeneous catalysts (acid, basic and Cu-based catalysts) on wood fractionation and light chemicals formation was investigated deeply in SC ethanol. In SC ethanol, catalysts favor light liquid products such as esters at the expense of biooil with a total oxygenates yield of 33 wt% relative to carbohydrates over β zeolite. However, we show that depending on the catalysts’ nature, wood components fractionation was completely changed, and this is particularly true with solid acid catalysts which promote cellulose deconstruction and the formation of solid carbon products. It is proposed that liquid products’ accumulation in the autoclave, in particular water, is at the origin of the wood fractionation changes which preclude its control by the choice of the SC organic solvent and conditions. Moreover, all the catalysts underwent severe leaching, which also contributed to the wood component fractionation changes.

Deconstructing sugarcane bagasse lignocellulose by acid-based deep eutectic solvents to enhance enzymatic digestibility

Biorefinery with deep eutectic solvent (DES) is an emerging processing technology to overcome the shortcomings of conventional biomass pretreatments. This work evaluates the biorefinery of sugarcane bagasse (SCB) with DES formulated with choline chloride as hydrogen bond acceptor and three hydrogen bond donors: lactic acid, citric acid, and acetic acid. Acetic acid showed unique ionic properties responsible for the selective removal of lignin and the deconstruction of cellulose to improve the digestibility of up to 97.61 % of glucan and 63.95 % of xylan during enzymatic hydrolysis. In addition, the structural characteristics of the polysaccharide-rich material (PRM) were analyzed by X-rays, ATR-FTIR, SEM, and enzymatic hydrolysis, and compared with the original material sample, for a comprehensive understanding of biomass deconstruction using different hydrogen bond donors (HBD) as DES pretreatment.

Deconstruction of pineapple peel cellulose based on Fe2+ assisted cold plasma pretreatment for cellulose nanofibrils preparation

An effective method based on Fe2+ assisted cold plasma (CP) to pretreat pineapple peel cellulose for the preparation of cellulose nanofibrils (CNF) was proposed in this study. The mechanism underlying Fe2+ assisted CP pretreatment based on the structural changes of cellulose was investigated. Results showed that the cellulose preabsorbed Fe2+, followed by 60 min of CP treatment, and then was nanofibrillated into CNF via ultrasonication. Fe2+ assisted CP treatment exhibited strong capacity to significantly decrease the degree of polymerization of cellulose and to enhance electrostatic repulsion of fibrils. Furthermore, the roughness and breakage were observed on the surface of pretreated cellulose. The content of inter-molecular hydrogen bonds and average crystallite size (Dhkl) also decreased significantly. The current results provided useful fundamental insights into Fe2+ assisted CP and cellulose interaction, which should be beneficial to developing new pretreatment methods for disintegrating cellulose into CNF.

Multiscale Molecular Simulation Strategies for Understanding the Delignification Mechanism of Biomass in Cyrene

In recent years, the cellulose-derived solvent Cyrene has piqued considerable interest in the green chemistry community despite only recently being available in the quantities required for solvent applications. Deconstruction of cellulose is an essential step in the production of fuel and value-added chemicals from lignocellulosic biomass. However, the high recalcitrance and heterogeneity of lignin hinder this process, necessitating the need to solubilize lignin. To understand the dissolution of lignin in Cyrene and Cyrene–cosolvent systems, multiscale molecular simulation approaches have been employed. Initially, the conductor-like screening model for real solvent (COSMO-RS) model was used to assess the thermodynamic properties of lignin in Cyrene and Cyrene–cosolvent systems. From the COSMO-RS calculations, the correlation between the predicted activity coefficient and the experimental lignin solubility was excellent. Further, classical molecular dynamics (MD) simulations were performed to evaluate the delignification of biomass by predicting structural and dynamic properties of lignin–solvent systems. The microscopic properties such as interaction energies, radius of gyration, solvent-accessible surface area, radial and spatial distribution functions (RDFs/SDFs), and hydrogen bonds were assessed to characterize lignin dissolution in these solvent mixtures and were validated with experimental data. From the MD simulations, it was observed that lignin adopts a coil-like structure in Cyrene and Cyrene:water mixtures, thereby dissolving the lignin, while lignin adopts a collapsed-like structure in the presence of water. The occupancy density of Cyrene is highly surrounded by the aryl and hydroxyl groups of lignin polymer rather than by water. The interaction energies between lignin and Cyrene and Cyrene–cosolvent were much stronger than that between lignin and water, explaining the higher biomass delignification in Cyrene-based solvents.

Production of high-yield 5-hydroxymethylfurfural from crystalline cellulose via one-pot conversion in molten salt hydrate/acetone and separation

5-hydroxymethylfurfural (5-HMF) is a crucial biomss-derived fuel precursor with versatile conversion pathways to advanced fuels. However, direct production of 5-HMF from crystalline cellulose is critical challenging due to the inefficient deconstruction of recalcitrant cellulose structure. Molten salt hydrates (MSHs) are unique in cellulose deconstruction and dissolution. In this work, a biphasic system of LiBr hydrate/acetone was established. It was found that 93.4% yield of 5-HMF can be efficiently converted from fructose without any catalysts. With the addition of AlCl3 to adjust Lewis acidity, an enhanced 5-HMF yield of 75.1% together with 13.7% of glucose were directly produced from cellulose, showing a promising carbon balance of 88.8%. A proposed mechanism for cellulose conversion was studied by density functional theory (DFT) calculations, demonstrating that the isomerization of glucose to fructose rather than crystalline cellulose hydrolysis was the rate-determining step. After reaction, the formed 5-HMF can be efficiently recycled from solvent via hyper-cross-linked polymer adsorption and the reaction system can be recycled for more than 5 times without reducing 5-HMF selectivity.

The Potential for Cellulose Deconstruction in Fungal Genomes

Fungal cellulolytic enzymes are carbohydrate active enzymes (CAzymes) essential for the deconstruction of the plant cell wall. Cellulolytic activity is described in some glycoside hydrolases (GH-cellulases) and in auxiliary activities (AA-cellulases) families. Across environments, these enzymes are mostly produced by some fungi and some bacteria. Cellulolytic fungi secrete these enzymes to deconstruct polysaccharides into simple and easy to metabolize oligo- and mono-saccharides. The fungal ability to degrade cellulose result from their repertoire of CAZymes-encoding genes targeting many substrates (e.g., xylan, arabinose). Over the past decade, the increased number of sequenced fungal genomes allowed the sequence-based identification of many new CAZyme-encoding genes. Together, the predicted cellulolytic enzymes constitute the fungal potential for cellulose deconstruction. As not all fungi have the same genetic makeup, identifying the potential for cellulose deconstruction across different lineages can help identify the various fungal strategies to access and degrade cellulose (conserved vs. variable genomic features) and highlight the evolution of cellulase-encoding genes. Here, the potential for cellulose deconstruction identified across publicly accessible, and published, fungal genomes is discussed.

Thermo-mechano-chemical deconstruction of cellulose for cellulose nanocrystal production by reactive processing

The commercial production of cellulose nanocrystals (CNCs) requires high concentration of sulfuric or other acids such as hydrochloric, phosphoric, and nitric acids. However, these acids and the involved process are corrosive, toxic, energy-intensive, and not environmentally safe. In this work, a batch mixer reactive process that entails high shear was implemented using 1-butyl-3-methylimidazolium chloride (BmimCl) media and molten oxalic acid dihydrate (OA) to produce CNCs from cellulose. Through this, a maximum CNC yield (59 wt%) was obtained with a mixture composition of 1:0.7:0.075 (Cellulose:BmimCl:OA, w/w/w) and a processing time of 2.5 min. Further investigation revealed that the particle size, degree of crystallinity, and thermal stability of the produced CNCs were found to be competitive with those of a commercial CNC product. This study asserts the potential industrial application of an efficient ionic liquid and molten organic acid treatment for CNC production via reactive processing in a batch mixer.

A cleaner processing approach for cellulose reinforced thermoplastic polyurethane nanocomposites

AbstractIn a manner of addressing challenges in scalable processing of thermoplastic polyurethane (TPU) nanocomposites through extrusion methods, this study reports a very clean processing approach of incorporating cellulose nanocrystal (CNC) into a TPU matrix, with no acid or organic‐solvents usage. It involves a mechanical deconstruction of microcrystalline cellulose (MCC) into nanoscale particles in water and polyol through scalable bead‐milling, vacuum drying, and followed by twin‐screw reactive extrusion with isocyanate and chain extender. The thermal stability of CNC was higher than that of typically acid‐hydrolyzed CNC and suitable for processing with the precursors of TPU at typical processing temperature range (175–190°C). The CNC incorporation at very low loadings (0.5, 0.8 wt%) through this methodology resulted in substantial enhancements in tensile properties (for example, up to 28% in strength and toughness) without any significant stiffening effect. Moreover, the nanocomposites retained elastic properties, including elongation at break (%), resilience, and creep resistance. Their chemical properties and thermal transitions were also found to support the retained thermoplastic behavior while improving mechanical performance.

Sustainable, superfast deconstruction of natural cellulosic aggregates toward intrinsically green, multifunctional gel

Natural cellulosic fibers/aggregates, being the most abundant eco-friendly resource, are a promising material solution toward the carbon neutrality and sustainability of our society. Further functionalization is required for their value-added applications but often involves complicated operations, high cost or toxic chemicals. Herein, we designed a simple and cost-effective strategy to enable super-fast yet completed deconstruction of cellulosic aggregates to cellulose molecules via microwave (MW)-assisted-ionic liquid (IL) engineering. MW radiation triggers the high-frequency swing of ions of IL, which leads to superfast heating behavior with a heating rate of ∼ 20 °C/s; meanwhile, the violent rotation of IL ions physically attacks cellulosic aggregates, accelerating their deconstruction. Our MW-IL strategy enables a superfast and complete deconstruction of cellulose (e.g., ∼240 s) compared with the conventional technology (>2400 s with non-complete deconstruction). The deconstructed cellulose can be facilely developed into the intrinsically green, multifunctional gel with high transparent, ion-conductive, injectable and reused advantages, which detects human motions accurately and continuously. The MW-IL-triggered deconstruction of cellulose opens a promising direction for the super-fast, cost-effectively preparation of high-quality cellulose materials, which are versatile toward advanced applications, including flexible electronics, micro/nano fluidics, and energy storage and conversion.

Roles of Ball Milling Pretreatment and Titanyl Sulfate in the Synthesis of 5-Hydroxymethylfurfural from Cellulose

Production of 5-hydroxymethylfurfural (HMF) from cellulose has been expected for a long time, but most catalytic systems generally give low yield and selectivity due to the difficulty of balancing cellulose deconstruction with the uncontrollable degradation of the target product under harsh conditions. Here we show that the ball milling pretreatment could markedly facilitate the conversion of microcrystalline cellulose to HMF by titanyl sulfate (TiOSO4). The ball milling pretreatment remarkably decreases the degree of crystallinity of microcrystalline cellulose with a reduction of molecular weight via disrupting the hydrogen bond and partially breaking the β-1,4-glycosidic bond. TiOSO4 functions as both Lewis and Brønsted acid to catalyze the efficient conversion of cellulose with high HMF yield (45.4%), far exceeding that (14.1%) from untreated cellulose. The reaction pathway was revealed by liquid chromatograph–mass spectra analysis. In addition, phosphorylated titanium dioxide also afforded a notable HMF yield (21.8%), showcasing the great potential of Ti-based heterogeneous catalyst.

Biorefinery aspects for cost-effective production of nanocellulose and high value-added biocomposites

Biomass waste can originate from a range of sectors, including agricultural, forestry, aquatic, and industrial waste. In order to establish a cost-effective biorefinery technology for industrializing nanocellulose-derived biomaterials, optimization of various factors affecting bioprocess is necessary. This article presents an overview of various cost-effective and greener enzymatic preparation strategies, mechanism of cellulose deconstruction and reconstruction, as well as the factors affecting bioprocesses is presented. It also discusses the role of endoglucanase, cellubiohydrolase and lytic polysaccharide monooxygenases in the enzymatic hydrolysis of process of cellulose. The potential applications of nanocellulose biocomposites in various promising fields are further presented and discussed in detail.

Processive Enzymes Kept on a Leash: How Cellulase Activity in Multienzyme Complexes Directs Nanoscale Deconstruction of Cellulose.

Biological deconstruction of polymer materials gains efficiency from the spatiotemporally coordinated action of enzymes with synergetic function in polymer chain depolymerization. To perpetuate enzyme synergy on a solid substrate undergoing deconstruction, the overall attack must alternate between focusing the individual enzymes locally and dissipating them again to other surface sites. Natural cellulases working as multienzyme complexes assembled on a scaffold protein (the cellulosome) maximize the effect of local concentration yet restrain the dispersion of individual enzymes. Here, with evidence from real-time atomic force microscopy to track nanoscale deconstruction of single cellulose fibers, we show that the cellulosome forces the fiber degradation into the transversal direction, to produce smaller fragments from multiple local attacks (“cuts”). Noncomplexed enzymes, as in fungal cellulases or obtained by dissociating the cellulosome, release the confining force so that fiber degradation proceeds laterally, observed as directed ablation of surface fibrils and leading to whole fiber “thinning”. Processive cellulases that are enabled to freely disperse evoke the lateral degradation and determine its efficiency. Our results suggest that among natural cellulases, the dispersed enzymes are more generally and globally effective in depolymerization, while the cellulosome represents a specialized, fiber-fragmenting machinery.

Assembling mini-xylanosomes with Clostridium thermocellum XynA, and their properties in lignocellulose deconstruction

Lignocellulose is a prominent source of carbohydrates to be used in biorefineries. One of the main challenges associated with its use is the low yields obtained during enzymatic hydrolysis, as well as the high cost associate with enzyme acquisition. Despite the great attention in using the fraction composed by hexoses, nowadays, there is a growing interest in enzymatic blends to deconstruct the pentose-rich fraction. Among the organisms studied as a source of enzymes to lignocellulose deconstruction, the anaerobic bacterium Clostridium thermocellum stands out. Most of the remarkable performance of C. thermocellum in degrading cellulose is related to its capacity to assemble enzymes into well-organized enzymatic complexes, cellulosomes. A mini-version of a cellulosome was designed in the present study, using the xylanase XynA and the N-terminus portion of scaffolding protein, mCipA, harboring one CBM3 and two cohesin I domains. The formed mini-xylanosome displayed maximum activity between 60 and 70 °C in a pH range from 6 to 8. Although biochemical properties of complexed/non-complexed enzymes were similar, the formed xylanosome displayed higher hydrolysis at 60 and 70 °C for alkali-treated sugarcane bagasse. Lignocellulose deconstruction using fungal secretome and the mini-xylanosome resulted in higher d-glucose yield, and the addition of the mCipA scaffolding protein enhanced cellulose deconstruction when coupled with fungal enzymes. Results obtained in this study demonstrated that the assembling of xylanases into mini-xylanosomes could improve sugarcane deconstruction, and the mCipA protein can work as a cellulose degradation enhancer.

How Do Shipworms Eat Wood? Screening Shipworm Gill Symbiont Genomes for Lignin-Modifying Enzymes.

Shipworms are ecologically and economically important mollusks that feed on woody plant material (lignocellulosic biomass) in marine environments. Digestion occurs in a specialized cecum, reported to be virtually sterile and lacking resident gut microbiota. Wood-degrading CAZymes are produced both endogenously and by gill endosymbiotic bacteria, with extracellular enzymes from the latter being transported to the gut. Previous research has predominantly focused on how these animals process the cellulose component of woody plant material, neglecting the breakdown of lignin – a tough, aromatic polymer which blocks access to the holocellulose components of wood. Enzymatic or non-enzymatic modification and depolymerization of lignin has been shown to be required in other wood-degrading biological systems as a precursor to cellulose deconstruction. We investigated the genomes of five shipworm gill bacterial symbionts obtained from the Joint Genome Institute Integrated Microbial Genomes and Microbiomes Expert Review for the production of lignin-modifying enzymes, or ligninases. The genomes were searched for putative ligninases using the Joint Genome Institute’s Function Profile tool and blastp analyses. The resulting proteins were then modeled using SWISS-MODEL. Although each bacterial genome possessed at least four predicted ligninases, the percent identities and protein models were of low quality and were unreliable. Prior research demonstrates limited endogenous ability of shipworms to modify lignin at the chemical/molecular level. Similarly, our results reveal that shipworm bacterial gill-symbiont enzymes are unlikely to play a role in lignin modification during lignocellulose digestion in the shipworm gut. This suggests that our understanding of how these keystone organisms digest and process lignocellulose is incomplete, and further research into non-enzymatic and/or other unknown mechanisms for lignin modification is required.

Unraveling a Lignocellulose-Decomposing Bacterial Consortium from Soil Associated with Dry Sugarcane Straw by Genomic-Centered Metagenomics.

Second-generation biofuel production is in high demand, but lignocellulosic biomass’ complexity impairs its use due to the vast diversity of enzymes necessary to execute the complete saccharification. In nature, lignocellulose can be rapidly deconstructed due to the division of biochemical labor effectuated in bacterial communities. Here, we analyzed the lignocellulolytic potential of a bacterial consortium obtained from soil and dry straw leftover from a sugarcane milling plant. This consortium was cultivated for 20 weeks in aerobic conditions using sugarcane bagasse as a sole carbon source. Scanning electron microscopy and chemical analyses registered modification of the sugarcane fiber’s appearance and biochemical composition, indicating that this consortium can deconstruct cellulose and hemicellulose but no lignin. A total of 52 metagenome-assembled genomes from eight bacterial classes (Actinobacteria, Alphaproteobacteria, Bacilli, Bacteroidia, Cytophagia, Gammaproteobacteria, Oligoflexia, and Thermoleophilia) were recovered from the consortium, in which ~46% of species showed no relevant modification in their abundance during the 20 weeks of cultivation, suggesting a mostly stable consortium. Their CAZymes repertoire indicated that many of the most abundant species are known to deconstruct lignin (e.g., Chryseobacterium) and carry sequences related to hemicellulose and cellulose deconstruction (e.g., Chitinophaga, Niastella, Niabella, and Siphonobacter). Taken together, our results unraveled the bacterial diversity, enzymatic potential, and effectiveness of this lignocellulose-decomposing bacterial consortium.