The aim of this work was to prepare a substrate for growing edible and medicinal mushrooms on soft-wood materials. A liquid part of digestate from biogas station was successfully used in spruce sawdust fermentation and Pleurotus ostreatus, Pleurotus eryngii, and Ganoderma lucidum were grown on the obtained substrates showing mycelial growth rates similar to control beech sawdust with values ranging from 4.1 to 5.54 mm/day. A six week-fermentation was demonstrated sufficient for removal of volatile extractives from the spruce sawdust (76 % removal efficiency) which elevated content was shown to be most critical for fungal growth. Simultaneously, a 47 % removal of resinous compounds and a decrease in C:N ratio of the growth substrate were observed in the sawdust fermentation in the presence of the liquid digestate. Among the ligninolytic enzymes, the produced growth substrates favored laccase production by tested fungi. From all of that it follows that the utilization of wastes from biogas production for the reuse of softwood wastes could make an environmental-friendly and economically viable biotechnology for mushroom production.