The genome of Schizophyllum commune encodes a diverse repertoire of degradative enzymes for plant cell wall breakdown. Recent comparative genomics study suggests that this wood decayer likely has a mode of biodegradation distinct from the well-established white-rot/brown-rot models. However, much about the extracellular enzyme system secreted by S. commune during lignocellulose deconstruction remains unknown and the underlying mechanism is poorly understood. In this study, extracellular proteins of S. commune colonizing Jerusalem artichoke stalk were analyzed and compared with those of two white-rot fungi Phanerochaete chrysosporium and Ceriporiopsis subvermispora and a brown-rot fungus Gloeophyllum trabeum. Under solid-state fermentation (SSF) conditions, S. commune displayed considerably higher levels of hydrolytic enzyme activities in comparison with those of P. chrysosporium, C. subvermispora and G. trabeum. During biodegradation process, this fungus modified the lignin polymer in a way which was consistent with a hydroxyl radical attack, similar to that of G. trabeum. The crude enzyme cocktail derived from S. commune demonstrated superior performance over a commercial enzyme preparation from Trichoderma longibrachiatum in the hydrolysis of pretreated lignocellulosic biomass at low enzyme loadings. Secretomic analysis revealed that compared with three other fungi, this species produced a higher diversity of carbohydrate-degrading enzymes, especially hemicellulases and pectinases acting on polysaccharide backbones and side chains, and a larger set of enzymes potentially supporting the generation of hydroxyl radicals. In addition, multiple non-hydrolytic proteins implicated in enhancing polysaccharide accessibility were identified in the S. commune secretome, including lytic polysaccharide monooxygenases (LPMOs) and expansin-like proteins. Plant lignocellulose degradation by S. commune involves a hydroxyl radical-mediated mechanism for lignocellulose modification in parallel with the synergistic system of various polysaccharide-degrading enzymes. Furthermore, the complex enzyme system of S. commune holds significant potential for application in biomass saccharification. These discoveries will help unveil the diversity of natural lignocellulose-degrading mechanisms, and advance the design of more efficient enzyme mixtures for the deconstruction of lignocellulosic feedstocks.

Microbial consortia represent promising candidates for aiding in the development of plant biomass conversion strategies for biofuel production. However, the interaction between different community members and the dynamics of enzyme complements during the lignocellulose deconstruction process remain poorly understood. We present here a comprehensive study on the community structure and enzyme systems of a lignocellulolytic microbial consortium EMSD5 during growth on corn stover, using metagenome sequencing in combination with quantitative metaproteomics. The taxonomic affiliation of the metagenomic data showed that EMSD5 was primarily composed of members from the phyla Proteobacteria, Firmicutes and Bacteroidetes. The carbohydrate-active enzyme (CAZyme) annotation revealed that representatives of Firmicutes encoded a broad array of enzymes responsible for hemicellulose and cellulose deconstruction. Extracellular metaproteome analysis further pinpointed the specific role and synergistic interaction of Firmicutes populations in plant polysaccharide breakdown. In particular, a wide range of xylan degradation-related enzymes, including xylanases, β-xylosidases, α-l-arabinofuranosidases, α-glucuronidases and acetyl xylan esterases, were secreted by diverse members from Firmicutes during growth on corn stover. Using label-free quantitative proteomics, we identified the differential secretion pattern of a core subset of enzymes, including xylanases and cellulases with multiple carbohydrate-binding modules (CBMs). In addition, analysis of the coordinate expression patterns indicated that transport proteins and hypothetical proteins may play a role in bacteria processing lignocellulose. Moreover, enzyme preparation from EMSD5 demonstrated synergistic activities in the hydrolysis of pretreated corn stover by commercial cellulases from Trichoderma reesei. These results demonstrate that the corn stover-adapted microbial consortium EMSD5 harbors a variety of lignocellulolytic anaerobic bacteria and degradative enzymes, especially those implicated in hemicellulose decomposition. The data in this study highlight the pivotal role and cooperative relationship of Firmicutes members in the biodegradation of plant lignocellulose by EMSD5. The differential expression patterns of enzymes reveal the strategy of sequential lignocellulose deconstruction by EMSD5. Our findings provide insights into the mechanism by which consortium members orchestrate their array of enzymes to degrade complex lignocellulosic biomass.

Livestock wastewater reuse could be a potential source for the distribution of antibiotics, antibiotic resistance bacteria (ARB), and antibiotic resistance genes (ARGs) in agricultural soil. In this study, soil samples were collected from different depths (0–60 cm) of farmland that has been subjected to long-term application of livestock wastewater. The vertical distribution of antibiotics, bacterial communities, and ARGs were assessed to identify the driving factors that could potentially influence the distribution of ARB and ARGs. The results demonstrated distinguished distributions of antibiotics along the soil depths, with tetracyclines (TCs) mainly found in the top 10 cm of the soil (0.11–0.31 μg/kg), while quinolones (QNs), sulfonamides (SAs), and macrolides (MLs) were detected in all 60 cm of soil depth (0.01–0.22 μg/kg). The selection pressure of antibiotics to microorganisms led to the proliferation of ARB, especially tetracycline-resistant bacteria and erythromycin-resistant bacteria. In terms of the distribution/abundance of ARGs, novA and tetA (58) were relatively higher in 0–10 cm surface soil, while vanRM and vanRF were mainly detected in the deeper soil. Different ARGs may have the same host bacteria, which lead to the emergence of multidrug resistant bacteria, such as Ilumatobacter sp., Aggregatilinea sp., Rhabdothermincola sp., and Ornithinimicrobium sp. Soil pH, electrical conductivity (EC), and moisture content (MC) could affect the distribution and proliferation of ARB and were found negatively correlated with most of the ARGs except macB. Therefore, it is potentially possible to eliminate/inhibit the spread of ARGs by adjusting these soil parameters. These findings provide insights into the distribution and dissemination of antibiotics, ARB, and ARGs in agricultural practices of livestock wastewater irrigation and provide effective mitigation strategies to ensure the safe use of livestock wastewater in agriculture.

The overuse of antibiotics has resulted in the prevalence of antibiotic resistance genes (ARGs) and antibiotic resistance bacteria (ARB) in the environment. High-density livestock farming is one of the major industries for antibiotic overuse. In this study, we sampled wastewater and manure at different stages of the feces treatment process from a dairy farm, as well as the soil in the farmland where the treated wastewater was being used for irrigation purpose. High-throughput Illumina sequencing was used to analyze the profiles of bacteria communities and ARGs. The results showed that the main ARG types were multidrug, aminoglycoside, glycopeptide, and tetracycline resistance genes, and Actinobacteria, Proteobacteria and Firmicutes were the main host bacteria phyla of these ARGs. The genus Nocardioides sp. and Ornithinimicrobium sp. were closely associated with the ARGs in the investigated samples. The relative abundances of ARGs in wastewater and manure were reduced by 68.5% and 62.1%, respectively, by the existing feces treatment process. Anaerobic fermentation and high-temperature fermentation were the most efficient treatment steps; the relative abundances of ARGs were reduced by 29.3% and 33.6% in the treated wastewater and manure, respectively. Irrigation with the treated wastewater significantly increased the abundance and diversity of ARGs and ARB in the surface soil of the farmland. The residual ARGs were found to transit through vertical gene transfer (VGT) and horizontal gene transfer (HGT) in soil. Therefore, the direct application of this inadequately treated wastewater and/or manure could risk spreading ARGs into the environment, and potentially impact human health. In order to effectively restrain the spread of ARGs, it is necessary to modify the wastewater and manure treatment processes and improve the regulations and guidelines of applying treated wastewater for irrigation.

Bacterial degradation mechanism for high chlorinated pentachlorobiphenyl (PentaCB) with worse biodegradability has not been fully elucidated, which could limit the full remediation of environments afflicted by the complex pollution of polychlorinated biphenyls (PCBs). In this research, a new PentaCB-degrading bacterium Microbacterium paraoxydans that has not been reported was obtained using enzymatic screening method. The characteristics of its intracellular enzymes, proteome and metabolome variation during PentaCB degradation were investigated systematically compared to non-PentaCB conditions. The findings indicate that the degradation rate of PentaCB (1 mg/L) could reach 23.9% within 4 hours and achieve complete degradation within 12 hours, with the mixture of intracellular enzymes being most effective at a pH of 6.0. During the biodegradation of PentaCB, the 12 up-regulated proteins characterized included ABC transporter PentaCB-binding protein, translocase protein TatA, and signal peptidase I (SPase I), indicating the presence of functional proteins for PentaCB degradation in both the cytoplasm and the outer surface of the cytoplasmic membrane. Furthermore, five differentially enriched metabolites were strongly associated with the aforementioned proteins, especially the up-regulated 1, 2, 4-benzenetriol which feeds into multiple degradation pathways of benzoate, chlorocyclohexane, chlorobenzene and aminobenzoate. These relevant results help to understand and speculate the complex mechanisms regarding PentaCB degradation by M. paraoxydans, which have both theoretical and practical implications for PCB bioremediation.

Facile and sensitive methods for detecting neonicotinoids (NEOs) in aquatic environments are crucial because they are found in extremely low concentrations in complex matrices. Herein, nitrogen-based magnetic conjugated microporous polymers (Fe₃O₄@N-CMP) with quaternary ammonium groups were synthesized for efficient magnetic solid-phase extraction (MSPE) of NEOs from tap water, rainwater, and lake water. Fe₃O₄@N-CMP possessed a suitable specific surface area, extended π-conjugated system, and numerous cationic groups. These properties endow Fe₃O₄@N-CMP with superior extraction efficiency toward NEOs. The excellent adsorption capacity of Fe₃O₄@N-CMP toward NEOs was attributed to its π–π stacking, Lewis acid–base, and electrostatic interactions. The proposed MSPE-HPLC-DAD approach based on Fe₃O₄@N-CMP exhibited a wide linear range (0.1–200 µg/L), low detection limits (0.3–0.5 µg/L), satisfactory precision, and acceptable reproducibility under optimal conditions. In addition, the established method was effectively utilized for the analysis of NEOs in tap water, rainwater, and lake water. Excellent recoveries of NEOs at three spiked levels were in the range of 70.4 to 122.7%, with RSDs less than 10%. This study provides a reliable pretreatment method for monitoring NEOs in environmental water samples.