This study aimed to optimize total nitrogen (TN) content in the medium and the growth temperature to improve the shelf life of Pleurotus eryngii Keneutari No. 2. We investigated the growth characteristics and changes in the O2/CO2 content in polypropylene film and performed sensory evaluations during cold storage under 1.5%, 1.8%, 2.1%, and 2.4% TN and at temperatures of 9°C, 12°C, and 15°C. The diameter and weight of the fruit bodies tended to increase with increasing nitrogen content, regardless of the growth temperature, and the highest weight per bottle (149.5 g) was obtained at 12oC and 2.4% TN. Regardless of the nitrogen content, leaf color deepened with decreasing temperature. The gas concentration in the film, which changes depending on storage time, was maintained for the longest period at 9oC and 2.1% TN. The sensory evaluation revealed no difference between the different nitrogen conditions tested, and the odor, discoloration, and decay were similar at 15oC and 9oC until day 28 of storage.