According to the nutrient requirement of the main oil recovery functional bacteria in the sandstone reservoir of Lu 9 well block，by-products of fermentation and agricultural wastes were selected as the main component of the nutrient medium，which also contained some auxiliary microbial growth factors. The optimum carbon source，nitrogen source and phosphorus source were selected through single factor experiment. The concentration of each component was preliminarily determined. On this basis，the significant influence factors of each component were analyzed by Plackett-Burman experiment and factor removal experiment，then the concentration of each component was determined. Finally，response surface test was used to further improve the concentration of each component. Moreover，oil displacement experiment was performed under simulated reservoir condition. The results showed that the nutrient system was efficient and cheap. The nutrient medium contained 0.15% by-products of fermentation and agriculture， 0.6% mineral salt with nitrogen content of 16.47%，and 0.25% mineral salt with nitrogen content of 26.16%. The oil emulsifying effect of the system was good. The copy number of hydrocarbon oxidation gene，which could activate the main oil recovery function gene，reached 10⁷ copies/mL. In the physical simulation oil displacement experiment，the recovery factor could increase 11.65 percentage points after injecting 0.4 PV nutrient system and air at a gas-liquid volume ratio of 8∶1. Compared with the formula used in the field，the optimum formula could reduce the cost by 20.25%. The screening and evaluation process of activation system could provide reference for the screening and evaluation of nutritional system.