Fig. 2

H₂O₂ accumulation in reactions with various electron donors in conjunction with TthLPMO9G. The figure illustrates the concentrations of accumulated H₂O₂, apparent over a 1-h period from reactions with 30 µM of a freshly prepared electron donor and 0.65 µM LPMO. Concentrations were measured at 10 min intervals. In the bar chart, the light blue bars represent the H₂O₂ concentration measured at 30 min, while the dark blue bars correspond to the H₂O₂ concentration measured at 60 min. The labels specify if the enzyme was present in the reactions, and the samples are arranged in an ascending sequence. Reaction mixtures, prepared as per the guidelines of the Amplex® Red Hydrogen Peroxide/Peroxidase Assay Kit, included specific concentrations of buffer, temperature, HRP, and Amplex® Red. Error bars represent standard deviations among duplicate experiments, each conducted at least twice. Throughout the entire experimental setup, the concentrations of the standards—0.5 μM H₂O₂, 3 μM H₂O₂, 5 μM H₂O₂, and 10 μM H₂O₂—remained stable