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Material, microbial and metabolism pathway perspectives on understanding the nitrogen/iron co-doped biochar enhanced methane production in brewery wastewater anaerobic digestion

Junguo He,Yili Wang,Yuanyi Zhao,Heliang Pang,Lin Li,Yunlong Liu,Xinxin Cui,Tong Wei,Suiyin Yu

At the present stage, the incorporation of modified biochar materials into anaerobic digestion (AD) systems has emerged as a pivotal strategy for the treatment of high-concentration organic wastewater and the promotion of methane production. This study proposes a novel methodology for a simple one-pot method, utilizing deep eutectic solvents (DES, urea/FeCl₃) to modify the preparation of nitrogen/iron co-doped straw biochar (DES BC), and it was applied for the first time with the objective of enhancing methane production in brewery wastewater AD. The incorporation of 1 g/L DES 1:1, 1:2, 1:3 BC into mesophilic AD systems resulted in increase the maximum methane production by a factor of 1.31, 2.84, and 1.50, respectively, in comparison with the control group that without DES BC. The enhancement mechanisms are attributed to three key aspects: (1) DES BC accelerated substrate conversion, as evidenced by rapid volatile fatty acid (VFA) consumption and microbial activity promotion, facilitated by its high specific surface area and pore volume providing attachment sites for functional microorganisms (e.g., Georgenia and Anaerolinea ). (2) the redox-active functional groups (─ OH, Fe O, N O) on DES BC enhanced electron exchange capacity (EEC), electron donating capacity (EDC), and electron accepting capacity (EAC), directly improving electron transfer efficiency, and (3) DES BC potentially promoted direct interspecies electron transfer (DIET) between syntrophic bacteria (e.g., Geobacteraceae , Desulfomicrobium ) and methanogenic archaea (e.g., Methanobacterium , Methanothrix ), optimizing methane yield without solely relying on microbial abundance. Metagenomic analysis further confirmed that DES BC increased the relative abundances of key enzymes in hydrogenotrophic and acetate decarboxylation methanogenesis pathways. This work provides innovative strategies for modifying biochar to advance brewery wastewater treatment and energy recovery.