Anti‑restriction protein KlcAHS enhances carbapenem resistance.

Abstract

The KlcAHS gene was previously identified as coexisting with the blaKPC‑2 gene in the backbone region of a series of blaKPC‑2‑harboring plasmids. The purpose of the present study was to determine the association between the KlcAHS and blaKPC‑2 genes. KlcAHS deletion and complementation experiments were used to evaluate the association between KlcAHS and carbapenem minimal inhibition concentrations (MICs). Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis was used to detect changes in the expression levels of blaKPC‑2 upon knocking out the KlcAHS gene in a blaKPC‑2‑harboring plasmid. The imipenem MIC of the transformants harboring ΔKlcAHSpHS10842 was lower (16µg/ml) than that of the transformants harboring wild‑type pHS10842 (32µg/ml), whereas the kanamycin MIC of the transformants harboring pET24a was lower (1,024µg/ml) than that of the transformants harboring pET24a‑KlcAHS (2,048µg/ml). The imipenem MICs of the two NM1049 Escherichia coli strains carrying plasmids pHS092839 or ΔKlcAHSpHS092839 exceeded 16µg/ml, whereas the ertapenem MIC of the host strains harboring ΔKlcAHSpHS092839 was 4µg/ml compared with ≥8 µg/ml observed in the host strains carrying pHS092839. The RT‑qPCR results demonstrated that the messenger RNA expression levels of blaKPC‑2 in the transformants carrying ΔKlcAHSpHS092839 were significantly downregulated (P=0.007) compared with those in the transformants carrying pHS092839. These findings revealed that KlcAHS elevated the MIC values of various antibiotics by upregulating the expression levels of blaKPC‑2. Therefore, KlcAHS can confer increased resistance to carbapenems in host strains. The survival probability of clinical pathogens may be enhanced by the presence of the KlcAHS gene in antibiotics used on a large scale.