Abstract: The combination effects of biochar and organic fertilizer on soil enzyme activities, microbial biomass carbon and nitrogen were studied to reduce the barriers of roductivity and establish a reasonable fertilization system on yellow-cinnamon soil. Under the condition of equal nitrogen, filed experiments were designed as: no fertilizer (CK), biochar (B), NPK fertilizers (NPK), biochar + NPK fertilizers (NPK+B), organic fertilizer + NPK fertilizers (NPK+M), biochar + organic fertilizer + NPK fertilizers (NPK+B+M). The soil enzyme activities, the microbial biomass carbon (MBC), the microbial biomass nitrogen (MBN) and some properties of soil, yield of wheat were measured by chloroform fumigation- K2SO4 lixiviating method and conventional analysis, respectively. The results showed that both biochar and organic fertilizer increased soil urease activity and enhanced microbial biomass carbon and nitrogen. The treatment NPK+M+B was the best of the all. Organic fertilizer improved β-glucosidase activity while biochar inhibited β-glucosidase activity. The MBC, MBN, soil urease activity and β- glucosidase activity were positively related to the yield significantly. The chemical fertilizer application combined with biochar and organic fertilizer is an optimal way to improve soil enzyme activities in the research.