Fifty-one isolates of Klebsiella pneumoniae were recovered from different hospitals in Northern Palestine, during September-December 2015. Results showed that the prevalence of ESBLs, MBLs and AmpC β-lactamases using multiplex PCR technique were 92.2%, 9.8 and 3.92%, respectively. For ESBL genes, TEM gene was the most dominant (72.5%) among K. pneumoniae isolates. Other genes were (0.0%), (17.6%) and (31.4%) for CTX-M, SHV and OXA genes, respectively. For AmpC β-lactamase producing K. pneumoniae, only DHA gene was detected and all MBL producing isolates were carried NDM gene. Molecular analysis by construction phylogenetic tree showed that all sequenced TEM, SHV, OXA, NDM and DHA genes were belonged to TEM-1, SHV-1, OXA-1, NDM-1 and DHA-1, respectively. In addition, 51%of K. pneumoniae harbored only class 1 integrons, whereas other tested classes were not detected. All class 1 integrons were detected in isolates carried β-lactamase genes. ERIC-PCR profile showed that these isolates were diverse and unrelated clones. This is the first study documented the prevalence and molecular characterization of β-lactamases producing K. pneumoniae isolates in Palestine Our results showed high occurrence of β-lactamases among K. pneumoniae isolates in Palestine. Based on these results we recommend the continuous monitoring and surveillance of the prevalence, proper control and prevention practices and effective antibiotic use will restrict the further spread of β-lactamases producing K. pneumoniae isolates within hospitals in this country.