Pseudomonas sp. Y-5, a strain with simultaneous nitrification and denitrification (SND) capacity, was isolated from the Wuhan Municipal Sewage Treatment Plant. This strain could rapidly remove high concentrations of inorganic nitrogen. Specifically, Pseudomonas sp. Y-5 removed 103 mg/L of NH4+-N in 24 h without nitrate or nitrite accumulation when NH4+-N was its sole nitrogen source. The NH4+-N removal efficiency (RE) was 97.26%, and the average removal rate (RR) was 4.30 mg/L/h. Strain Y-5 also removed NO3--N and NO2--N even in aerobic conditions, with average RRs of 4.39 and 4.23 mg/L/h, respectively, and REs of up to 99.34% and 95.81% within 24 h. When cultured in SND medium (SNDM-1), strain Y-5 achieved an NH4+-N RE of up to 97.80% and a total nitrogen (TN) RE of 93.01%, whereas NO3--N was fully depleted in 48 h. Interestingly, high nitrite concentrations did not inhibit the nitrification capacity of Y-5 when grown in SNDM-2, the RE of NH4+-N and TN reached 96.29% and 94.26%, respectively, and nitrite was consumed completely. Strain Y-5 also adapted well to high concentrations of ammonia (~401.68 mg NH4+-N/L) or organic nitrogen (~315.12 mg TN/L). Our results suggested that Pseudomonas sp. Y-5 achieved efficient simultaneous nitrification and denitrification, thus demonstrating its potential applicability in the treatment of nitrogen-polluted wastewater.