Unveiling molecular DOM reactomics and transformation coupled with multifunctional nanocomposites under anaerobic conditions: Tracking potential metabolomics and pathways

Anaerobic digestion (AD) offers great potential for pollutant removal and bioenergy recovery. However, it faces challenges when using livestock manure (LSM) as a feedstock given its high content of refractory materials (e.g., lignocellulose, long-chain carbohydrates, lipids, and crude protein). This would significantly inhibit AD-microbial activities, reduce organic transformation efficiency and limit gas production. To overcome this, multifunctional metal-doped hydrochars (HCs) were introduced here as AD supplements/accelerators, given that LSM degradation under AD results in complex dissolved organic matter (DOM). To assess this, the current study investigates the molecular interactions/transformations within DOM during LSM-AD coupled with metal-doped HCs, via batch-mode experiments. Expansive data mining techniques were employed to analyze DOM using Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Substantial increments in peptide-like along with decrements in highly unsaturated-like molecules were observed in HC@MnCl₂ containing-system. This indicates an increased capability for substrate hydrolysis and potential utilization of soluble microbial products (SMPs) (i.e., highly unsaturated-like molecules), leading to enhanced methane recovery (223.23 mL/g-VS_added, 1.77 times more than the control). However, accumulation of DOM-highly unsaturated molecules (i.e., a lack of SMPs' degradation) accompanied with low methane production (39.68 mL/g-VS_added) was noticed for HC@NiFe₂O₄. DOM reactivity during LSM-AD was validated via paired mass difference molecular network, indicating predominance of CHO and N-containing groups’ transformations for HC@MnCl₂ and HC@NiFe₂O₄, respectively. Potential metabolites and abundant pathways were verified via KEGG database. This study improves our understanding of LSM-AD-DOM complex transformation matrix, the fate of bioavailable/recalcitrant compounds, and identification of potential DOM regulators from thousands of molecules.