Versatile peroxidase from white rot fungus Bjerkandera adusta was over-expressed in a soluble form in Escherichia coli. In the constructed enzyme model based on the selected gene from B. adusta, the active sites for oxidation of Mn(2+) ions and for oxidation of aromatic substrates were identified, both characteristic for versatile peroxidase. For over-expression of the recombinant enzyme different host strains, media formulations, growth temperatures, and fusion partners were tested. With the bacterial strain BL21(DE3)pLysS cultivated at 25 degrees C in auto-induction medium and presence of heme, a soluble peroxidase with incorporated heme and activity against different substrates was obtained. By exploiting an appropriate expression system and providing suitable culture conditions, the recombinant fungal peroxidases in soluble form can be produced in bacteria.