Given the numerous environmental problems caused by chemical fertilizer overuse, agricultural practices are shifting toward the development of environmentally friendly nitrogen (N) fertilizers. In this study, a pot experiment was conducted to investigate the effect of poly(gamma-glutamic acid) (gamma-PGA), an effective fertilizer synergist, on rapeseed (Brassica napus cv. Qinyou No. 9) productivity, soil N pools, and soil enzymes. Seven treatments were implemented: T-1, a check without urea; T-2, a check with urea only; T-3, urea mixed with glutamic acid; T-4, urea mixed with 3 mg of gamma-PGA per kilogram of soil; T-5, urea mixed with 10 mg of gamma-PGA per kilogram of soil; T-6, urea coated with gamma-PGA (0.9%, m/m); and T-7, urea coated with gamma-PGA (3.1%, m/m). The results demonstrated that the application of gamma-PGA increased the above-ground fresh weight of rapeseed by 7.57-9.26% and the grain yield per pot by 6.46-10.98% compared with T-2. The rapeseed grain yield significantly increased from 15.94 +/- 0.55 g pot(-1) in T-2 to 17.69 +/- 0.78 g pot(-1) in T-7. With the increase in pod numbers and seed numbers per pod, the seed yield was significantly improved after gamma-PGA application. Culturable soil microbial count and microbial diversity were increased by gamma-PGA application. In addition, the effects of application method of gamma-PGA have a more important role on the soil microflora than its amount. The reduced loss of N can be attributed to the capacity of gamma-PGA to increase the "NH4+ turnover pool." More mineral N in the soil was immobilized by clays, soil microbes, and gamma-PGA at the early growth stage, and the immobilized N was then released at the later growth stage for plant growth. Moreover, urease activity also increased with gamma-PGA application. These findings indicated that the urea coated with gamma-PGA could be a better choice for agricultural applications.