Contribution of the Fenton reaction and ligninolytic enzymes to soil organic matter mineralisation under anoxic conditions

Highlights: • Fenton reaction and oxidative enzymes explain high CO₂ efflux from anaerobic soils. • Synergism of Fenton reaction and lignin peroxidase activity induce high CO₂ efflux. • H₂¹⁸O₂ labelling allows identifying and differentiating abiotic oxidative processes. Mechanisms of carbon dioxide (CO₂) release from soil in the absence of oxygen were studied considering the Fenton process, which encompasses the reaction of H₂O₂ with Fe(II) yielding a hydroxyl radical (OH), in combination with manganese peroxidase (MnP) and lignin peroxidase (LiP). This study aimed to explain the high rate of soil organic matter (SOM) mineralisation and CO₂ release from humid temperate rainforest soils under oxygen-limited conditions. The investigated mechanisms challenge the traditional view that SOM mineralisation in rainforest is slow due to anaerobic (micro)environments under high precipitation and explain intensive CO₂ release even under oxygen limitation. We hypothesised that the Fenton reaction (FR) greatly contributes to the CO₂ released from SOM mineralised under anaerobic conditions especially in the presence of ligninolytic enzymes. We used a novel technique that combines labelled H₂¹⁸O₂ and Fe(II) to induce the FR and measured CO¹⁸O, Fe(II) solubilisation, and peroxide consumption in a closed gas circulation system for 6 h. Maximal CO₂ amount was released when the FR was induced in combination with LiP addition. The CO₂ efflux with LiP was 10-fold that of abiotic FR reactions without enzymes, or in soils amended with MnP. This was consistent with i) the contribution of ¹⁸O from peroxide to CO₂ release, ii) peroxide consumption, and iii) Fe(II) solubilisation by FR. The amount of consumed peroxide was closely correlated with the CO¹⁸O derived from soil without enzyme addition or with LiP addition. Concluding, abiotic Fenton Reaction coupled with oxidative enzymes, such as LiP, are crucial for SOM oxidation under anaerobic conditions, e.g. in temperate rainforest soils.