Crude oil as a non-renewable resource is creating new challenges in many industrial sectors. Unsteady costs of crude oil at present and expected increases in the future are due to its limited availability as a finite resource, and these costs negatively impact the industry for wood-based panels, which use petrochemical resins in binding agents. Furthermore, wood panels that are conventionally bonded using urea formaldehyde diffuse formaldehyde into the surrounding air. To achieve independence from petrochemical products and harmful formaldehyde emissions, alternatives for their substitution are in demand. An alternative approach is the enzymatic activation of lignin located on the surface of thermomechanical pulp (TMP) fibers. The present study shows the results of internal bond strength (DIN EN 319 1993), modulus of rupture (DIN EN 310 1993), and thickness swelling (EN 317 2003) of medium-density fiberboards (MDF) bonded with laccase-mediator-system (LMS). Caffeic acid (CA), 4-hydoxy benzoic acid (HBA), and vanillic alcohol (VAl) were used as mediators. The physical and technological properties of MDF, such as internal bond strength, modulus of rupture, and thickness swelling, mostly fulfilled the European standards.