Background: Enterobacter spp. has been reported as an important opportunistic, multi resistant bacterial pathogen and incriminated in nosocomial infections. Objectives: This study aimed to determine the prevalence of Enterobacter spp. & their antibiotic susceptibility profiles and to detect the ability of Enterobacter spp. to produce siderophore-mediated strategy for iron acquisition. Methodology: Enterobacter spp. were identified and confirmed by standard microbiological methods and Vitek- 2 system. Their Antibiotic susceptibility profiles were determined by the modified Kirby Bauer disk diffusion method. Also, extended-spectrum β-lactamases (ESβL) and metallo-β-lactamase production were tested by combined disc diffusion method. Irp2 Virulence gene was detected by conventional polymerase chain reaction (PCR). Results: A total of 50 Enterobacter isolates were collected in this study from different samples from hospital (Group1), community acquired infected patients (Group 2) and from the feces of healthy volunteers (Group 3). E.cloacae was the most predominant enterobacter species (54.0%). The highest isolation rate of Enterobacter spp. was from sputum samples. About 75%, 70% and 62.5% were ESβL producers, and about 62.5%, 50% and 37.5% were MβL producers among the three studied groups. Irp2 gene was detected in 65.5% of HAI group, 40% of CAI group and 25% of carriers. Conclusion: The combination between multi-drug resistance and siderophores' virulence genes in Enterobacter species is worrying, since prevalence of these opportunistic pathogens causing nosocomial infections is increasing.