Abstract The kEC value (=extractable part of microbial biomass C) of the fumigation-extraction (FE) method was assessed on the basis of 153 soils (94 arable, 46 grassland and 13 forest soils) by indirect calibration using the fumigation-incubation (FI) method. Sixty-six soils were investigated for the first time and the data on a further 87 soils were obtained from the literature. The single kEC values ranged from 0.23 to 0.84. A split according to the form of land use resulted in a significantly (Scheffe, P = 0.05) lower kEC value for the arable soils (0.42; n = 94) in comparison to those for the grassland (0.49; n = 46) and the forest soils (0.51; n = 13). This difference is mainly due to the significant effects of the respiration rate measured in non-fumigated control samples of the FI method which was used for calibration of the kEC value. For that reason, I investigated the effects of incubation temperature (22°, 25° and 28°C) on biomass C data obtained by the FI method, and thus on the kEC value of the FE method, and discuss further problems of direct and indirect calibration. Based on experimental and literature data, I conclude that the kEC values of Vance et al. (Soil Biology & Biochemistry19, 703–707, 1987) and Wu et al. (Soil Biology & Biochemistry22, 1167–1169, 1990) remain valid. A kEC value of 0.38 can be recommended for C analysis by dichromate consumption and a kEC value of 0.45 for that by UV-persulfate or oven oxidation.