The enzymatic hydrolysis of lignocellulosic polymers is generally considered the rate-limiting step to methane production in anaerobic digestion of lignocellulosic biomass. The present study aimed to investigate how the hydrolytic microbial communities of three different types of anaerobic digesters adapted to lignocellulose-rich wheat straw in continuous stirred tank reactors operated for 134 days. Cellulase and xylanase activities were monitored weekly using fluorescently-labeled model substrates and the enzymatic profiles were correlated with changes in microbial community compositions based on 16S rRNA gene amplicon sequencing to identify key species involved in lignocellulose degradation. The enzymatic activity profiles and microbial community changes revealed reactor-specific adaption of phylogenetically different hydrolytic communities. The enzymatic activities correlated significantly with changes in specific taxonomic groups, including representatives of Ruminiclostridium, Caldicoprobacter, Ruminofilibacter, Ruminococcaceae, Treponema, and Clostridia order MBA03, all of which have been linked to cellulolytic and xylanolytic activity in the literature. By identifying microorganisms with similar development as the cellulase and xylanase activities, the proposed correlation method constitutes a promising approach for deciphering essential cellulolytic and xylanolytic microbial groups for anaerobic digestion of lignocellulosic biomass.

Carbon capture and utilization (CCU) covers an array of technologies for valorizing carbon dioxide (CO₂). To date, most mature CCU technology conducted with capture agents operates against the CO₂ gradient to desorb CO₂ from capture agents, exhibiting high energy penalties and thermal degradation due to the requirement for thermal swings. This Perspective presents a concept of Bio-Integrated Carbon Capture and Utilization (BICCU), which utilizes methanogens for integrated release and conversion of CO₂ captured with capture agents. BICCU hereby substitutes the energy-intensive desorption with microbial conversion of captured CO₂ by the methanogenic CO₂-reduction pathway, utilizing green hydrogen to generate non-fossil methane.

Anaerobic digestion (AD) is a process in which microorganisms, under oxygen-free conditions, convert organic matter into biogas and digestate. Normally, only 40–70% of biomass is converted into biogas; therefore, digestate still contains significant amounts of degradable organic matter and biogas potential. The recovery of this residual biogas potential could optimize substrate utilization and lower methane emissions during digestate storage and handling. Post-treatment methods have been studied with the aim of enhancing the recovery of biogas from digestate. This review summarizes the studies in which these methods have been applied to agricultural and wastewater digestate and gives a detailed overview of the existing scientific knowledge in the field. The current studies have shown large variation in outcomes, which reflects differences in treatment conditions and digestate compositions. While studies involving biological post-treatment of digestate are still limited, mechanical methods have been relatively more explored. In some cases, they could increase methane yields of digestate; however, the extra gain in methane has often not covered treatment energy inputs. Thermal and chemical methods have been studied the most and have yielded some promising results. Despite all the research conducted in the area, several knowledge gaps still should be addressed. For a more thorough insight of the pros and cons within post-treatment, more research where the effects of the treatments are tested in continuous AD systems, along with detailed economic analysis, should be performed.

Biogas production from organic waste is a waste-to-energy technology with the potential to contribute significantly to sustainable energy production. Upgrading of biogas using in situ biomethanation with hydrogen has the potential for surplus electricity storage, and delivery of biogas with a methane content of >90%, allowing for easier integration into the natural gas grid, as well as conversion to other products. Microbial communities in biomethanation reactors undergo changes, however, these changes are largely unexplored. In the present study, metagenome-resolved protein stable isotope probing (Protein-SIP) was applied to laboratory scale batch incubations operating under anaerobic digestion, and (pre-adapted) biomethanation conditions, fed with ¹³C-labelled bicarbonate, in order to gain insight into the microbial activities during CO₂-reduction. The strongest and most microbially diverse isotopic incorporation was observed in the pre-adapted biomethanation incubation. Furthermore, divergent incorporation of ¹³C-labelled bicarbonate was also observed in the Wood-Ljungdahl pathway, with the anaerobic digester incubations primarily showing labelled proteins in the peripheral pathways leading toward production of energy and biomass. The pre-adapted biomethanation incubations consumed H₂ and CO₂, but did not convert it to CH₄, suggesting the production of acetate in these incubations, which was supported by heavy labelling of key enzymes in the Wood-Ljungdahl pathway. Twelve (ten high quality) metagenome-assembled genomes (MAGs) coding for ¹³C-incorporated proteins were extracted from the metagenome, eight of which contained one or more of the key genes in the Wood-Ljungdahl pathway, one of which was affiliated to Methanosarcina. Together, the findings in the present study deepen our knowledge surrounding microbial communities in biomethanation systems, and contribute to the development of better strategies for implementation of biogas upgrading and microbial management.