Klebsiella pneumoniae carbapenemase (KPC)-encoding genes containing promoter-deletions (blaKPC-2a, blaKPC-2b, and blaKPC-2c) have disseminated in Enterobacteriaceae. The minimal inhibitory concentrations (MICs) to β-lactams in clinical KPC-producing Enterobacteriaceae range from susceptible to high-level resistant, resulting in diagnostic problems. To better understand the variability in β-lactam MICs among KPC-producing Enterobacteriaceae, three isoforms of blaKPC-2 gene were used to transform Escherichia coli W4573 and its deletion mutant of an efflux pump (AcrAB) to examine the effects on β-lactam susceptibility. MICs to β-lactams in E. coli W4573 and its acrAB mutant strain increased 1- to 500-fold (MIC from 0.125 to 64 μg mL−1 of aztreonam) in the blaKPC-2a, blaKPC-2b, and blaKPC-2c transformants compared with the cloning vector alone. However, transformants of the acrAB mutant strain remained susceptible to all β-lactams tested except for aztreonam and carbenicillin. Levels of the three promoters’ length and carbapenemase activities in the transformants harboring the blaKPC-2a, blaKPC-2b, and blaKPC-2c were correlated to the levels of β-lactam MICs in both E. coli W4573 and its mutant of an efflux pump (AcrAB). Overall, these results suggest that promoter-deletions of blaKPC-2 gene and AcrAB may be associated with the variability in β-lactam MICs in KPC-producing Enterobacteriaceae.