Background: Nitrogen(N) deposition alters litter decomposition and soil carbon(C) sequestration by influencing the microbial community and its enzyme activity. Natural atmospheric N deposition comprises of inorganic N(IN) and organic N(ON) compounds. However, most studies have focused on IN and its effect on soil C cycling, whereas the effect of ON on microbial enzyme activity is poorly understood. Here we studied the effects of different forms of externally supplied N on soil enzyme activities related to decomposition in a temperate steppe. Ammonium nitrate was chosen as IN source, whereas urea and glycine were chosen as ON sources. Different ratios of IN to ON(Control, 10:0, 7:3, 5:5, 3:7, and 0:10) were mixed with equal total amounts of N and then used to fertilize the grassland soils for 6 years.Results: Our results show that IN deposition inhibited lignin-degrading enzyme activity, such as phenol oxidase(POX) and peroxidase(PER), which may restrain decomposition and thus induce accumulation of recalcitrant organic C in grassland soils. By contrast, deposition of ON and mixed ON and IN enhanced most of the C-degrading enzyme activities, which may promote the organic matter decomposition in grassland soils. In addition, the β-N-acetylglucosaminidase(NAG) activity was remarkably stimulated by fertilization with both IN and ON, maybe because of the elevated N availability and the lack of N limitation after long-term N fertilization at the grassland site. Meanwhile, differences in soil p H, soil dissolved organic carbon(DOC), and microbial biomass partially explained the differential effects on soil enzyme activity under different forms of N treatments.Conclusions: Our results emphasize the importance of organic N deposition in controlling soil processes, which are regulated by microbial enzyme activities, and may consequently change the ecological effect of N deposition. Thus, more ON deposition may promote the decomposition of soil organic matter thus converting C sequestration in grassland soils into a C source.