This study was conducted to investigate the effect of culture temperature on the growth rate and protease activity of Antarctic microorganisms. The Antarctic microorganisms PAMC 25641, 25614, 25719 and 25617 were obtained from the Polar and Alpine Microbial Collection (PAMC) at the Korea Polar Research Institute. These microorganisms were confirmed for the excretion of protease on a plate with skim milk. The identification of microorganisms was carried out using the 16S rDNA sequencing method. PAMC 25641 showed the highest protease activity among the subjects tested, and PAMC 25617 exhibited the highest growth rate. The growth rates of the microorganisms were not affected by temperature, except for PAMC 25617. However, protease activities were increased for all strains in a temperature dependent fashion. These results suggest the possible application of Antarctic microorganisms for the efficient production of low temperature proteases.