Biochar Mitigates N₂O Emission of Microbial Denitrification through Modulating Carbon Metabolism and Allocation of Reducing Power

To elucidate the direct effects of biochar on denitrification metabolism at the cellular level, the global response of model denitrifying soil bacterium (Paracoccus denitrificans) to biochar addition was investigated by physiological, proteomic, and metabolomics analyses. The enhancement effect on denitrification was positively correlated with its pyrolysis temperatures (300-500 °C) and dosages (0.1-1%), regardless of precursors [corn straw (CS) or wheat straw). Moreover, the stimulating effect of CS biochar made at 500 °C (CS-500) was mainly attributed to the bulk particles rather than the released soluble compounds. Without direct contact with cells, bulk CS-500 particles might directly modulate the carbon metabolism by the adsorption of extracellular metabolites. Since carbon flux to storage was shifted to oxidative catabolism and growth assimilation, more share of the produced reducing power was used for nitrogen reduction. Meanwhile, except for nitrate reductase, both protein expressions and enzyme activities of nitrite reductase, nitric oxide reductase, and nitrous oxide reductase were up-regulated. Accordingly, the accumulation of N₂O was reduced by 98% due to the optimized electron distribution among denitrifying enzymes. Eventually, the growth rate of Pc. denitrificans enhanced because of the improved energy utilization efficiency. These results updated the regulation mechanism of biochar on denitrification metabolism and N₂O mitigation.