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An increase in the antibiotic resistance among members of theEnterobacteriaceaefamily has been observed worldwide. Multidrug-resistant Gram-negative rods are increasingly reported. The treatment of infections caused byEscherichia coliand otherEnterobacteriaceaehas become an important clinical problem associated with reduced therapeutic possibilities. Antimicrobial carbapenems are considered the last line of defense against multidrug-resistant Gram-negative bacteria. Unfortunately, an increase of carbapenem resistance due to the production ofKlebsiella pneumoniaecarbapenemase (KPC) enzymes has been observed. In this study we describe the ability ofE. colito produce carbapenemase enzymes based on the results of the combination disc assay with boronic acid performed according to guidelines established by the European Community on Antimicrobial Susceptibility Testing (EUCAST) and the biochemical Carba NP test. Moreover, we evaluated the presence of genes responsible for the production of carbapenemases (blaKPC,blaVIM,blaIMP,blaOXA-48) and genes encoding otherβ-lactamases (blaSHV,blaTEM,blaCTX-M) amongE. coliisolate. The tested isolate ofE. colithat possessed theblaKPC-3andblaTEM-34genes was identified. The tested strain exhibited susceptibility to colistin (0.38 μg/mL) and tigecycline (1 μg/mL). This is the first detection ofblaKPC-3in anE. coliST479 in Poland.