K. pneumoniae is one of the most isolated agents related to infectious diseases. The presence of multidrug resistant strains harboring both resistance and virulence genes in K. pneumoniae isolated from hospital settings, hospital sewage and aquatic environment, is considering a problem for clinician and a threat to Public Health around the world. This study aimed to identify phenotypic and genotypic profile of K. pneumoniae isolated from aquatic environment, hospital sewage and clinical isolates and their ability to adhere, invade and persist within human cell. A total of 30 MDR strains were isolated and presented at least one resistance gene for ESBL, PMQR, AME or carbapenem. MDR K. pneumoniae strains also presented virulence genes for type 1 and type 3 pili and iron uptake system. These isolates were able not only to form biofilm in different scales, but also to adhere, invade and persist in VERO cell line. Even though K. pneumoniae is considered one of the most important opportunistic pathogens, knowledge of the mechanism by which this bacterium causes different diseases is still unclear and studies needs to be done.