The objectives of the study were to investigate mineral soil profiles as a living space for microbial decomposers and the relation of microbial properties to soil acidity. We estimated microbial biomass C on concentration (μg g−1 DW) as well as on volume basis (g m−2) and the microbial biomass C to soil organic C ratio along a vertical gradient from L horizon to 20 cm in the mineral soil and along a gradient of increasing acidity at five beech forest stands in Germany. Microbial biomass C concentration ranged from 17,000–34,000 μg Cmic g−1 DW in the litter layer and decreased dramatically down the profile to 29–264 μg Cmic g−1 DW at 15–20 cm depth in the mineral soil. This represents depth gradients of microbial biomass C concentrations ranging from a factor of 65 in slightly acidic and up to 875 in acidic soils. In contrast, microbial biomass C calculated on a volume basis (g Cmic m−2) showed a different pattern since a considerable part of the microbial biomass C was located in the mineral soils. In the soil profile 22–34% of the microbial biomass C was found in the mineral soil at strictly acidic sites and as much as 64–88% in slightly acidic soils. The microbial biomass C to soil organic carbon ratios decreased in general down from the L horizon in the forest floor to 0–5 cm depth in the mineral soils. In strongly acidic mineral soils however, the C to soil organic carbon ratio increased with depth, suggesting a positive relation to increasing pH. We conclude from depth gradients of soil pH and microbial biomass C to soil organic carbon ratio that pH affects this ratio at acidic sites. The inter-site comparison indicates that acidity restricts microbial biomass C in the mineral soils.