Recently, with numerous environmental problems being caused by chemical fertilizer overuse, agricultural practices are shifting toward the development of environmentally friendly N fertilizers. In this study, pot and field experiments were simultaneously conducted to investigate the effect of poly(y-glutamic acid) (γ-PGA) on the yield, N use efficiency, and soil microenvironment of wheat. Our study demonstrates a statistically significant increase in winter wheat, number of tillers, seed number per spike, yield, soil microbial biomass N (SMBN), and soil enzymes after γ-PGA application. The highest grain yield of 7435.69 ± 55.91 kg ha-1 was obtained after γ-PGA application in the field experiment, which was 7.17% higher than the urea control. The N recovery efficiency increased by 11.81%-14.00% and 11.30%-11.38% after the application of γ-PGA in pot and field experiments, respectively. More mineral nitrogen in soil was immobilized by the microbes after γ-PGA application at the early growth stage of wheat. The immobilized nitrogen was gradually released at the late growth stage. The results demonstrate that γ-PGA can be used as a fertilizer synergist.