Microbial electrolysis cell-anaerobic digestion (MEC-AD) is an emerging technology for methane production. However, low substrate degradation efficiency remains a challenge when processing protein substrates. This study developed a MIL-100(Fe) carbon cloth anode to enhance methane production and substrate degradation in MEC-AD. The effects of MIL-100(Fe) prepared under hydrothermal (H-MIL-100(Fe)) and room temperature conditions (R-MIL-100(Fe)) were compared. Results indicated that H-MIL-100(Fe) and R-MIL-100(Fe) increased cumulative methane production by 16.01% and 14.99%, respectively compared to normal cloth, each influencing methane production through distinct mechanisms. Electrochemical characterization showed that H-MIL-100(Fe) enhanced the electrochemical performance more significantly due to the enrichment of Geotalea, with the oxidation current improved by 7.39-fold (R-MIL-100(Fe) increased it by only 2.95-fold) to promote growth of Methanobacterium. Metagenomic analysis revealed that R-MIL-100(Fe) tended to metabolize amino acids into methane rather than support cellular life activities, indicating its practicality under limited substrate concentration. In summary, R-MIL-100(Fe) shows greater potential for application due to its mild synthesis conditions and advantages in treating complex substrates.
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