Fig. 5

Recombinant expression of NcLPMO9C using P. pastoris followed by the hydrocoerulignone assay. LPMO expression was induced after 29 h using methanol. The fermentation medium was supplemented with copper sulfate to reach a final concentration of 100 µM. A sample was immediately taken after copper addition to the medium to obtain a reference. Green pentagons show wet biomass, blue squares show extracellular protein concentration, black diamonds show activity measured with 500 µM hydrocoerulignone as substrate (100 mM sodium acetate buffer, pH 6.0, 20 µl sample volume) and orange stars show activity with 1000 µM 2,6-DMP (50 mM sodium phosphate buffer, pH 7.5, 100 µl sample volume). The activity of 2,6-DMP is magnified in the inset, which has the same units as the X-axis and the right Y-axis. All activity measurements were performed for 300 s at 30 °C by the addition of 100 µM H₂O₂. All data are expressed as mean values (± SD) from three independent measurements