Microbiomes enrichment from a biogas plant

Lignocellulose biomasses (LCB), including spent mushroom substrate (SMS), pose environmental challenges if not managed properly, contributing to environmental pollution. However, these renewable resources hold immense potential for biofuels and chemicals production. With the mushroom market growth expected to amplify SMS quantities, repurposing or disposal strategies are critical. This study explores utilizing SMS for cultivating microbial communities to produce carbohydrate-active enzymes (CAZymes). Addressing a research gap in utilizing anaerobic digesters for enriching SMS-utilizing microbiomes, this study investigates microbial diversity and secreted CAZymes under varied temperatures (37C, 50C, and 70C) and substrates (SMS, carboxymethylcellulose, and xylan). Enriched microbiomes demonstrated temperature-dependent preferences for cellulose, hemicellulose, and lignin degradation, supported by thermal and elemental analyses. Enzyme assays confirmed lignocellulolytic enzyme secretion correlating with substrate degradation trends. Notably, thermogravimetric coupled with differential scanning calorimetry (TGA-DSC) emerged as a rapid approach for saccharification potential determination of LCB. Microbiomes isolated at mesophilic temperature secreted thermophilic hemicellulases exhibiting robust stability and superior enzymatic activity compared to commercial enzymes, aligning with biorefinery conditions. PCR-DGGE and metagenomic analyses showcased dynamic shifts in microbiome composition and functional potential based on environmental conditions, impacting CAZyme abundance and diversity. The meta-functional analysis emphasized the role of CAZymes in biomass transformation, indicating microbial strategies for lignocellulose degradation. Temperature and substrate specificity influenced the degradative potential, highlighting the complexity of environmental-microbial interactions. This study illuminates temperature-driven microbial selection for lignocellulose degradation, unveiling thermophilic xylanases with industrial promise. Insights gained contribute to optimizing enzyme production and formulating efficient biomass conversion strategies. Understanding microbial consortia responses to temperature and substrate variations elucidates bioconversion dynamics, emphasizing tailored strategies for harnessing their bio-technological potential.