Industrial utilization of lignin is of high interest since it represents around 30% of all nonfossil-based carbon sources worldwide. For various applications of lignosulfonates such as for dispersants or adhesives a larger molecular weight is essential. Here, we investigated laccase-catalyzed polymerization of lignosulfonate directly from the pulp and paper industry in the presence and absence of natural and synthetic mediators with and without oxygen supply. For example, laccase-mediated polymerization in the presence of a 2.5 mM TEMPO as mediator with a 10 cm 3 min −1 oxygen flow rate led to a 12-fold increase of the molecular weight, while without TEMPO a 13-fold increase was achieved. In contrast, without an external oxygen supply, only a 7-fold increase in molecular weight was achieved compared to a 4-fold increase for the TEMPO−laccase system. Fluorescence intensity, phenol content, and size exclusion chromatography measurements indicate that generally in the presence of high concentrations of mediators, such as TEMPO, vanillin, HBT, and 2,6-dimethoxyphenol, oxidation of other structural units in lignosulfonates may counteract desired polymerization reactions. In summary, for laccase-catalyzed polymerization of lignosulfonates, an external oxygen supply was found to be much more beneficial than the presence of laccase mediators.