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Additional file 1. Supplementary material. The genomes of the two most abundant species were independently assembled, their scaffolds were ordered by manual curation, and their closed chromosomes were drawn using Artemis [1]. The same software (parameters: windows size 10,000 bp and step size 200 bp) was also used to determine the GC content and the GC skew. The COG annotation was converted to “EMBL” format using in-house Perl scripts and added to the graphical representation in “gff” format reporting the gene positions predicted by Prodigal. COG codes assigned to the proteins were used to color the genes ( https://www.ncbi.nlm.nih.gov/research/cog ). The reconstruction of the five most abundant species metabolisms were also performed to investigate their role within the community. The reconstruction was performed from KEGG codes obtained using eggNOG mapper. The resulting networks were visually inspected in KEGG database using KEGG Mapper – Color Pathway, and results are depicted in Figures S5-S9. Table S1. Reactors pH, VFA, dissolved H2 concentration (H2l) and H2 transfer coefficient (kLa) under steady state conditions at period IV. Descriptive biochemical data of the analyzed phase were collected in a previous work and are reported here [2]. Table S2. Reactors' performances under steady state conditions at period 6. QOUT represents the total gas output rate [L/LR.d] and QOUT,T the total theoretical gas output rate [L/LR.d]. As described by Bassani et al. PCH4 indicates the CH4 production rate, YCH4 the CH4 yield, nH2 and nCO2 the hydrogen and CO2 conversion efficiency, respectively [2]. Figure S1. Representation of the reconstructed genome of Methanothermobacter wolfeii DTU-UNIPD957. Figure S2. Representation of the reconstructed genome of Lymnochordia sp. GSMM975. Figure S2. Representation of the reconstructed genome of Lymnochordia sp. GSMM975. Figure S4. Overall import and export fluxes (> 1 mmol/gDW/hr) of the community from the environment. Figure S5. Metabolic reconstruction of Acetomicrobium sp. GSSM972. Figure S6. Metabolic reconstruction of Methanothermobacter wolfeii GSSM957. Figure S7. Metabolic reconstruction of Firmicutes sp. GSSM974. Figure S8. Metabolic reconstruction of Firmicutes sp. GSSM966. Figure S9. Metabolic reconstruction of Limnochordia sp. GSSM975. Figure S10. Reactions and corresponding genetic evidence of CO2 reduction pathways included in the GSMM. Figure S11. FISH image of the microbial community.

Genome scale metabolic models of the five most abundant species present in the sampled biofilm and the result of model assessment with memote (v.0.13)

Statistics of filtered illumina reads and raw ONT-Nanopore reads optained with FastQC (v.0.11.9) and NanoPlot (v.1.32.1), respectively.