Yersinia enterocolitica causes various diseases in humans, including enteritis. The onset of such diseases is closely related with the expression of important virulence factors, particularly outer membrane proteins (OMPs). The expression of OMPs depends on several factors, including temperature, and origin, biotype and serotype of the bacteria. Recently, concerns over food safety have increased along with the demand for the development of sensitive, rapid, and pathogen-specific detection methods. To develop a suitable detection method for Y. enterocolitica isolated from Korean moutainspring water and pig feces, the OMP expression patterns were analyzed phenotypically and immunologically using 12 representative strains from 51 Y. enterocolitica Korean isolates. A 38-kDa OMP was commonly observed in all strains. However, additional OMPs were also observed in different biotypes and serotypes as well as bacterial origins, by incubating Y. enterocolitica at a low temperature. The specificity of the 38-kDa OMP was confirmed by a Western blot analysis with antisera against Y. enterocolitica and Brucella abortus. The results, therefore, indicate that the 38-kDa OMP could be used as a marker for detecting Y. enterocolitica in the environment or for seromonitoring.