Pan-genome analysis is used to interpret genome heterogeneity and diversification of bacterial species. Here, we present pan-genome analysis of 22 strains of Enterococcus mundtii. The GenBank file of E. mundtii strains that have been isolated from different sources i.e., human fecal matter, soil, leaf, dairy products, and insects was downloaded from National Center for Biotechnology Information (NCBI) database and analyzed using BPGA-1.3.0 (Bacterial Pan Genome Analysis) pipeline. Out of a total, 4503 gene families, 1843 belongs to the core genes whereas 1,762 gene families represent the accessory genes and 898 gene families depict the unique genes among all the selected genomes. Majority of the core genes belongs to the categories of Metabolism (37.83%) and Information storage & processing (29.84%) whereas unique genes belongs to the category of Information storage & processing (48.08%). Further, accessory genes are almost equally present in both functional categories i.e. Information storage & processing and Metabolism (34.34% and 32.27% respectively). Further, subset analysis on the basis of the origin of isolates exhibits presence and absence of exclusive gene families. The observation suggests that even closely related strains of a species show extensive disparity in genome owing to their ability to adapt to a specific environment.