Fallow deer are animals kept on large-scale pastures, which influence soil properties, including the content of nutrients in the soil and the flow of these nutrients in the soil–plant–animal system. Therefore, the aim of this case study was to analyze the variability in the macronutrient content and biochemical properties of soil under rotational grazing conditions of farmed fallow deer. Fallow deer grazed in two summer pens from April to November, and in the winter pen from December to March. The analyses included the determination of several soil parameters to capture sensitive soil changes and assess potential degradation of the soil environment in response to grazing: pHKCl, TOC, TN, N-NO3, N-NH4+, total (P, K, Mg, Ca, and Na) and available (Pav, Kav, Mgav) forms of macronutrients, and selected soil enzymes (dehydrogenase, acid phosphatase and alkaline phosphatase, and urease). The results showed that the pHKCl in the plots used by farm fallow deer was lower than in the soil of the control object. Moreover, the TN and TOC content in the soils of all pastures was statistically significantly lower than in the soil without grazing, while for TOC, the same effect was observed in the winter pasture soil. The content of N-NO3− was several times higher than that of N-NH4+ in the soils of the studied breeding plots. The relationships noted indicate that grazing is important in the transformation of soil organic matter, which is influenced by the relationship between carbon and nitrogen. In addition, the grazing of farmed fallow deer had a negative effect on the content of P in the soils of all pastures and a positive impact on the accumulation of K. Winter grazing had a positive effect on the amount of Pav and Kav in the soil, but a negative effect on the content of Mgav, compared to grazing in the summer quarters. In the plots used for pastures, the activity of soil enzymes was higher than in the control soil. It has been shown that the influence of fallow deer farming on the soil environment of pastures is not unequivocal. Therefore, long-term monitoring of changes in the properties of these soils is necessary.