Fig. 3

Apparent hydrogen peroxide production by two batches of AA10_07. The figure shows apparent H₂O₂ production by 3 µM AA10_07 in 50 mM sodium phosphate buffer, pH 6.0 supplied with 50 µM ascorbic acid, 5 U/ml HRP, 100 µM Amplex Red and 1% (v/v) DMSO. SEC−/SEC+ labels indicate the procedure used to remove unbound copper from the LPMO sample after copper saturation. SEC−, gravity-flow desalting column; SEC+, high-resolution SEC chromatography. Excess copper control reactions (grey bars) were set up using protein-free samples, obtained by ultrafiltration. These samples contained the same amount of free copper as the LPMO preparation used in the experiment. The two reactions shown to the left are control reactions: “background”, reaction without enzyme; “0.2 µM Cu(II)”, reaction without enzyme and with addition of 0.2 µM Cu(II). Error bars indicate standard deviations between triplicates