Climate warming is predicted to raise the mean global temperature by 1 °C in the next 50 years, and this change is believed to be capable of affecting soil organic matter cycling and nutrient availability. With the aim of increasing knowledge on the response of forest soils to the ongoing climate change, we used altitude as a proxy for temperature change and studied chemical and biochemical properties of European beech (Fagus sylvatica L.) forest soils at two altitudes (800 and 1000 m) from central Apennines (Italy). Results showed that 1 °C of mean annual air temperature difference between the sites at the two altitudes had greater effect on the mineral horizons than on the organic horizons. At higher altitude, mineral soil had limited development, higher pH, and higher organic matter content due to the lower efficiency of the microbial community. Enzymatic activities of the organic horizons were generally not affected by altitude. Conversely, we observed a higher activity of xylosidase, β-glucosidase, alkaline phosphomonoesterase, arylsulfatase, and leucine-aminopeptidase in the sub-superficial horizons (Bw1 and Bw2) of the soils at 1000 m. We hypothesized that, as a response to environmental and climatic constraints occurring at higher altitude, plant roots increase the production of enzymes directly and/or indirectly by triggering the microbial community through exudation.