In the food industry, ensuring food safety during transportation and storage is vital, with temperature regulation preventing spoilage. However, airborne contamination through foodborne pathogens remains a concern. Listeria monocytogenes, a psychrotolerant foodborne pathogen, has been linked to various foodborne outbreaks. Therefore, understanding how its airborne characteristics depend on the growth temperature is imperative. As a result, when the L. monocytogenes was floated in air for 30 and 60 min, the surviving population of 15 °C-grown L. monocytogenes that was suspended in air and attached on the surface was significantly higher than L. monocytogenes grown at 25°C and 37 °C. The fatty acid analysis revealed a significantly higher proportion of shorter chain fatty acids in L. monocytogenes grown at 15 °C compared to those grown at 37 °C. Under aerosolization, L. monocytogenes encountered osmotic and cold stresses regardless of their growth temperature. Transcriptomic analysis showed that stress response related genes, such as oxidative and cold stress response, as well as PTS system related genes were upregulated at 15 °C, resulting in the enhanced resistance to various stresses during aerosolization. These results provide insights into the different responses of aerosolized L. monocytogenes according to the different growth temperatures, highlighting a critical factor in preventing airborne cross-contamination.