The effects of γ-PGA (0.1%, 0.2%, 0.3% and 0.4%) application on soil agglomeration structure, soil nutrient balance, spinach yield and economic benefit were studied by pot experiment. The results showed that with the increasing of γ-PGA application rate, the water-stable aggregates (WR_0.25), the average weight diameter (MWD) and geometric mean diameter (GMD) increased, while the agglomerate destruction rate (PAD) and the fractal dimension (D) decreased, which indicated that γ-PGA had a significant effect on improving soil agglomeration structure. With the increasing of γ-PGA application rate, the absorption of nitrogen, phosphorus and potassium by spinach decreased, while soil nutrient residue and surplus amount increased, and the apparent loss of nutrients decreased. Among the five treatments, the soil nutrient balance condition of the 0.1% application treatment was the best, while the 0.4% application rate had a negative effect on crop nutrient absorption and soil nutrient balance. Spinach yield and economic benefit decreased significantly with the increasing of γ-PGA application rate, and the yield in 0.4% application treatment decreased by 42.98% compared with the control (field without adding γ-PGA). The fitting results of the spinach economic benefits showed that the profit of the treatment applicated 0~0.1% γ-PGA was higher than that of the treatment without γ-PGA. In summary, the 0.1% application rate was better for improving soil structure and nutrient balance, promoting crop yield and crop nutrient absorption, which provided a theoretical reference for the further application of γ-PGA in actual production.