Four biostimulants (BS): WCDSs, wheat condensed distiller solubles; PA-HE, hydrolyzed poultry feathers; CGHE, carob germ enzymatic extract; and RB, rice bran extract were applied annually at 4.7 t organic matter (OM) ha −1 for a 3-year period to a Xerollic Calciorthid soil to evaluate their efficiency in soil restoration. Their effects on the plant cover, soil enzymatic activities and the structure of the soil microbial community by analysing phospholipid fatty acids (PLFAs) were determined. Application of BS that contain higher amounts of protein and higher percentage of peptides under 3 kDa had a greater effect on the soil biological properties, possibly due to the low molecular weight protein content can be easily assimilated by soil microorganisms. Following 3 years of successive soil amendment, the dehydrogenase activity was 4.6, 9.6, and 17.6% higher in PA-HE-amended soils than in the RB, CGHE and WCDS-amended soils, respectively. The urease activity was 5.3, 14.5, and 28.8% higher in PA-HE-amended soils than in the RB, CGHE and WCDS-amended soils, respectively. The phosphatase activity was 8, 15.3, and 20.2% higher in PA-HE-amended soils than in the RB, CGHE and WCDS-amended soils, respectively. The arylsulfatase activity was 16, 21.1, and 27.2% higher in PA-HE-amended soils than in the RB, CGHE and WCDS-amended soils, respectively. Total soil phospholipid fatty acid (PLFA) concentration was significantly ( p < 0.05) higher in BS-amended soil than control soil. Principal component analysis discriminated between the BS-amended soils, mainly based on content of lower molecular weight peptides. Thus, PA-HE and RB were grouped and differentiated from CGHE and WCDS, respectively. After 3 years of treatment, vegetal cover was 11.4, 17.7, 24.1, and 85.8% higher in PA-HE-amended soils than in the RB, CGHE, WCDS treatments and control soil. These results suggested that under semiarid climatic conditions the application of BS with higher amounts of protein (>50%) and a higher percentage of peptides under 0.3 kDa (>60%) notably increased the soil enzymatic activities, induced changes in microbial community because the protein with lower molecular weight can be more easily absorbed by soil microorganisms, and also favoured the establishment of vegetation, which will protect the soil against erosion and will contribute to its restoration.