A thinned and an unthinned treatment were compared in a 62-year-old pine stand located in northeastern Germany (Brandenburg, Ost-Prignitz, Revier Beerenbusch) (year of thinning: 1999, degree of canopy opening: 0.4). Samples of the organic layer (O) and the mineral horizon (Aeh) of an acid brown earth were collected along a transect at each treatment in November 2003 and April 2004. Substrate induced respiration, basal respiration, and a suite of enzymes involved in the degradation of lignocellulose (endocellulase, exocellulase, β-glucosidase, endoxylanase, exoxylanase, phenoloxidase, peroxidase) were assayed. Microbial community structure and relative biomass of bacteria, actinomycetes, and fungi were assayed by phospholipid fatty acid analysis. Five years after thinning, microbial biomass, basal respiration, and enzyme activities in both soil layers did not differ significantly between thinned and unthinned treatments. However, the analysis of soil microbial community structure revealed a significant difference between the thinned and unthinned treatment at both sampling dates. Thus, it was concluded that thinning had not yet resulted in any response in soil microbial activities at the site under study, but since early evidence of change in the microbial community was detected, long-term monitoring and additional studies on mineralization activities are required.