Fig. 7
Comparative analysis of individual product release (in nC*min) from cellulose oxidation catalyzed by TthLPMO9G variants, WT (dark grey) and the S28A mutant (blue), following a 16-h reaction period. Each reaction incorporated a 0.1% PASC concentration, 4 μΜ LPMO, and 1 mM electron donors, and was diluted threefold with water prior to analysis via ion chromatography HPAEC-PAD. All experiments were carried out in 50 mM sodium acetate buffer, pH 6.0, at 45 °C. The chromatograms on the left delineate all eluted products, labeled ox-DPA1 to ox-DPA5 for the 13–19 min retention window. The bar chart on the right presents a comparative analysis of each eluted product for the WT and the S28A mutant under the influence of ascorbic acid and both gallic and caffeic acids. Control reactions without the enzyme addition consistently resulted in zero area. Product patterns ratios were derived from summed peak areas and normalized to the smallest value. Bars denote mean values, with error bars indicating the standard error derived from two independent experiments, each performed at least twice