Phosphorus (P) limitation will play a key role in the productivity of agriculture in the coming decades. Struvite is an ammonium magnesium phosphate mineral that can be recovered from wastewater-treatment plants and can be considered as an alternative source of P. However, the impact of struvite on the plant yield and, particularly, on the soil microbial community is barely known. Here, we tested the impacts of struvite, sewage sludge, and their combination on the barley yield, soil macro and micronutrients, and biochemical and microbiological soil properties. Amendment with struvite alone and its combination with sludge increased the availability of P in soil, the plant uptake of P and Mg, and the barley yield. The analysis of phospholipid fatty acids (PLFAs) and the utilization of metaproteomics approaches revealed significant effects of struvite on the biomass of Gram-positive bacteria and, particularly, on actinobacterial and verrucomicrobial populations in soil.