This study aimed to evaluate the use of softwood and hardwood waste for the production of levulinic acid by one-stage conversion using microwave radiation combined with acid catalysis. The analysis demonstrated that the type and concentration of the acid used, the concentration of biomass in the reaction mixture and pressure value had the greatest impact on the yield of levulinic acid. The highest efficiency of carbohydrate conversion to levulinic acid, regardless of the type of raw material, was achieved using a pressure of 225 PSI and sulfuric acid as a catalyst. Maximum yield from biomass, ca. 16.5% for cherry wood chips and ca. 25% for pine chips, was obtained using sulfuric acid at a concentration of 1% v/v and 2% v/v, respectively, for the following process parameters: Exposure time 20 min, biomass concentration 3.3%, and the pressure of 225 PSI. The ratio of actual yield to theoretical yield was high: 64.7% ± 4.5% for pine chips and 43.4% ± 1.0% for cherry wood chips. High efficiency of the presented method of biomass conversion to levulinic acid indicates the possibility of its use for waste management in the wood processing industry. High concentration of levulinic acid in the post-reaction mixture allows for cost-effective extraction and purification of the compound.