This study analyzed the effect of enzymatic aging (horseradish peroxidase) of biochars on their content of solvent extractable (Ctot) and freely dissolved (Cfree) polycyclic aromatic hydrocarbons (PAHs). Physicochemical properties and phytotoxicity of pristine and aged biochars were also compared. The study used biochars obtained at 500 or 700 °C from sewage sludges (SSLs) or willow. Compared to SSL-derived biochars, willow-derived biochars were more susceptible to enzymatic oxidation. Aging increased the specific surface area and pore volume of most SSL-derived biochars. An opposite direction, however, was found in the willow-derived biochars. Low-temperature biochars, regardless of their feedstock, underwent physical changes, such as removal of labile ash components or degradation of aromatic structures. The enzyme caused an increase in the content of Ctot light PAHs in biochars (by 34–3402 %) and heavy PAHs (≥4 rings) in the low-temperature SSL-derived biochars (by 46–713 %). In turn, the content of Cfree PAHs decreased in aged SSL-derived biochars (by 32–100 %). In the willow-derived biochars the bioavailability of acenaphthene increased (by 337–669 %), while the immobilization degree of some PAHs was lower (25–70 %) compared to the SSL-derived biochars (32–83 %). Nevertheless, aging positively affected the ecotoxicological properties of all biochars by increasing their stimulation effects or removing their phytotoxic effects on both Lepidium sativum seed germination and root growth. Significant relationships between the changes in Cfree PAH content, pH and salinity of SSL-derived biochars and seed germination/root growth inhibition were found. The study demonstrates that the risk associated with application of SSL-derived biochars, regardless of the type of SSL and pyrolysis temperature, can be lower in terms of Cfree PAHs than in the case of willow-derived biochars. Regarding to Ctot PAHs, high-temperature SSL-derived biochars are safer than low-temperature ones. In the case of application of high-temperature SSL-derived biochars, these with moderate alkalinity and salinity will not bring risks for plants.