Changes in soil enzyme activities and respiration were investigated in response to a series fertilization regimes in a long-term field experiment in order to evaluate nutrition equipoise in a fluvo-aquic soil at the Fengqiu National Agro-Ecological Experiment Station in Fengqiu, China under different fertilization treatments including control (CK), nitrogen + potassium (NK), phosphorus + potassium (PK), nitrogen + phosphorus (NP）, nitrogen + phosphorus + potassium (NPK), organic manure (OM), and 1/2OM +1/2 NPK. Results showed that soil enzyme activities (invertase, urease and phosphatase) and respiration were significantly influenced by long-term fertilization. In general, organic manure fertilization was more beneficial for improving the soil respiration and soil hydrolytase activities related with the recycling of carbon, nitrogen and phosphorus than chemical fertilizer application. Balanced chemical fertilizer application was more beneficial than unbalanced chemical fertilizer application except the treatment of nitrogen + phosphorus. It suggested that organic manure application will maintain or improve soil quality, especially to soil biochemical properties.