Soluble protein makes an important contribution to the release of carbon dioxide (CO2) in soils, however, knowledge on the respiration and C use efficiency (CUE) characteristics of soluble protein-derived C by soil microorganisms remains limited. To address this issue, we sampled surface soils (0−10 cm) from seven tree monocultures and investigated the temporal dynamics of turnover of 14C-labelled soluble protein-derived C by soil microorganisms. Two double first-order exponential kinetic decay models were applied to analyze the mineralization data (i.e., with and without abiotic protein-surface interactions). The model incorporating the immobilization of protein on the non-living solid phase exhibited the best fit to the experimental data (R2 > 99.6%). Our results suggest that 66.1−73.9% of the soluble protein-derived C was immobilized by the non-living solid phase in soils. After uptake by the soil microbial community, 8.0−13.8% of the C was rapidly respired as CO2, while 15.0−20.8% was used in anabolic processes, resulting in a CUE of 55.1−70.2%. However, there was little effect of forest type on protein turnover rate in the soil. The C:N ratio of soil microbial biomass (C/Nmic) was positively related to the CUE of protein and showed less variation within a forest type. Compared with soil microbial biomass C and N, C/Nmic could serve as a better indicator of the CUE of protein by soil microorganisms. This study sheds light on the respiration and CUE characteristics of soluble protein-derived C by soil microorganisms at afforested sites and enhances our understanding of the trade-off between the metabolism of protein-derived C by soil microorganisms and its immobilization by the non-living solid phase in soils.