The substrate availability for microbial biomass (MB) in soil is crucial for microbial biomass activity. Due to the fast microbial decomposition and the permanent production of easily available substrates in the rooted top soil mainly by plants during photosynthesis, easily available substrates make a very important contribution to many soil processes including soil organic matter turnover, microbial growth and maintenance, aggregate stabilization, CO2 efflux, etc. Naturally occurring concentrations of easily available substances are low, ranging from 0.1 mu M in soils free of roots and plant residues to 80 mM in root cells. We investigated the effect of adding C-14-labelled glucose at concentrations spanning the 6 orders of magnitude naturally occurring concentrations on glucose uptake and mineralization by microbial biomass. A positive correlation between the amount of added glucose and its portion mineralized to CO2 was observed: After 22 days, from 26% to 44% of the added 0.0009 to 257 mu g glucose C g(-1) soil was mineralized. The dependence of glucose mineralization on its amount can be described with two functions. Up to 2.6 mu g glucose C g(-1) soil (corresponds to 0.78% of initial microbial biomass C), glucose mineralization increased with the slope of 1.8% more mineralized glucose C per 1 mu g C added, accompanied by an increasing incorporation of glucose C into MB. An increased spatial contact between microorganisms and glucose molecules with increasing concentration may be responsible for this fast increase in mineralization rates (at glucose additions <2.6 mu g C g(-1)). At glucose additions higher than 2.6 mu g C g(-1) soil, however, the increase of the glucose mineralization per 1 mu g added glucose was much smaller as at additions below 2.6 mu g C g(-1) soil and was accompanied by decreasing portions of glucose C-14 incorporated into microbial biomass. This supports the hypothesis of decreasing efficiency of glucose utilization by MB in response to increased substrate availability in the range 2.6-257 mu g C g(-1) (=0.78-78% of microbial biomass Q. At low glucose amounts, it was mainly stored in a chloroform-labile microbial pool, but not readily mineralized to CO2. The addition of 257 mu g glucose C g(-1) soil (0.78 mu g C glucose mu g(-1) C micro-organ isms) caused a tag phase in mineralization of 19 h, indicating that glucose mineralization was not limited by the substrate availability but by the amount of MB which is typical for 2nd order kinetics. (C) 2008 Elsevier Ltd. All rights reserved.