Fig. 5

Dependence of the [H₂O₂]_t=0 and \(v_{{\left( {{\text{H}}_{2} {\text{O}}_{2} } \right)}}\) on pre-incubation time of LF at 50 °C. Reactions were made in 50 mM sodium acetate (pH 5.0) at 50 °C. The concentration of BMCC was 0.6 g L⁻¹ and that of LF was 10% (v/v). a Time curves of the formation of soluble products (expressed in glucose equivalents, Glc_eq) upon incubation of BMCC with TrLPMO9A in the presence of LF that was pre-incubated at 50 °C for different time (as defined on the plot). Shown are average values, and S.D, obtained from experiments with TrLPMO9A concentrations of 0.1 µM, 0.2 µM, or 0.5 µM (see Additional file 1: Fig. S5 for the individual curves). Solid lines show linear regression of the data according to Eq. 2. b, c Dependence of (b) the [H₂O₂]_t=0 and (c) the \(v_{{\left( {{\text{H}}_{2} {\text{O}}_{2} } \right)}}\), on pre-incubation time of LF at 50 °C. The values of [H₂O₂]_t=0 and \(v_{{\left( {{\text{H}}_{2} {\text{O}}_{2} } \right)}}\) were found by fitting the data in a to Eq. 2 and using a stoichiometry of 1.32 Glc_eq/H₂O₂. The values of [H₂O₂]_t=0 were also measured using an alternative approach (designated with SmLPMO10A on b) where LF was pre-incubated at 50 °C but LPMO reaction was done at 25 °C using the SmLPMO10/CNW system and short reaction times (Additional file 1: Fig S7). Solid lines in panel B show best-fits of the linear regression analysis. Error bars represent S.D. and are from three independent measurements each made at different concentration of TrLPMO9A (Additional file 1: Fig. S5)