The Influence of Biochar Pretreatment on Cell Immobilisation and Biochar Augmentation During Anaerobic Digestion of Cellulose

Biochar (BC) is widely utilised to enhance methane production from the anaerobic digestion (AD) process due to its physicochemical properties. This study investigates the influence of biochar pretreatment on its behaviour and performance in AD systems. Pre-treated biochar derived from different feedstocks via slow pyrolysis was applied in Biochemical Methane Potential (BMP) tests at a dosage of 0.25% (w/v), and its impact on both the methane yield and digestion kinetics during cellulose degradation was evaluated. Pre-treatment was performed via steam autoclaving, serving as both a sterilisation and surface-modification step aimed at improving the physicochemical characteristics of biochar, particularly its capacity for microbial immobilisation. The results demonstrated that the effect of autoclave pretreatment on methane production was strongly feedstock-dependent. Autoclaved rice husk biochar (AC-RH550) markedly inhibited methane production, whereas autoclaved softwood biochar (AC-SW550) exhibited the best performance among all biochars tested, achieving the highest methane yield of 382.2 (mL CH4 g−1 VS), corresponding to an increase of 18.3% compared to non-pretreated SW550 and 9.2% relative to the control. This improvement is attributed to the modifications in biochar surface properties, promoting improved microbial activity and biofilm formation potentially improving direct interspecies electron transfer (DIET). Additionally, biochar may act as a buffering agent and provide adsorption sites for inhibitory intermediates such as VFA. Cell immobilisation experiments further confirmed that autoclaving enhanced biochar porosity and hydrophilicity, encouraging better colonisation and biofilm formation. Scanning electron microscopy (SEM) and microbial analyses verified increased cell attachment on pretreated biochars. Overall, these findings highlight the critical role of pretreatment in optimising biochar functionality for anaerobic digestion applications and cell immobilisation.