GLO1 Contributes to the Drug Resistance of Escherichia coli Through Inducing PER Type of Extended-Spectrum β-Lactamases

Abstract

Background

Escherichia coli-associated antimicrobial resistance (AMR) issue so far needs urgent considerations. This study aims to screen the potent genes associated with extended-spectrum β-lactamases (ESBLs) in drug-resistant Escherichia coli and elucidate the specific drug-resistant mechanism.

Methods

Clinical ESBLs-EC samples were obtained based on the microbial identification, and the whole genome was sequenced. In combination with the significantly enriched pathways, several differently expressed genes were screened and verified by RT-PCR. Furthermore, through knocking out glyoxalase 1 (GLO1) gene and transfecting overexpressed plasmids, the potential relationship between GLO1 and ESBLs was then investigated. Lastly, the concentrations of β-lactamases in bacteria and supernatant from different groups were examined by enzyme-linked immunosorbent assay (ELISA).

Results

After successful isolation and identification of ESBLs-EC, the whole genome and eighteen differential metabolic pathways were analyzed to select differently expressed genes, including add, deoD, guaD, speG, GLO1, VNN1, etc. RT-PCR results showed that there were no differences in these genes between the standard bacteria and susceptible Escherichia coli. Remarkably, the relative levels of four genes including speG, Hdac10, GLO1 and Ppcdc were significantly increased in ESBLs-EC in comparison with susceptible strains, whereas other gene expression was decreased. Further experiments utilizing gene knockout and overexpression strains confirmed the role of GLO1. At last, a total of 10 subtypes of β-lactamases were studied using ELISA, including BES-, CTX-M1-, CTX-M2-, OXA1-, OXA2-, OXA10-, PER-, SHV-, TEM-, and VEB-ESBLs, and results demonstrated that GLO1 gene expression only affected PER-β-lactamases but had no effects on other β-lactamases.

Conclusion

SpeG, Hdac10, GLO1 and Ppcdc might be associated with the drug-resistant mechanism of Escherichia coli. Of note, this study firstly addressed the role of GLO1 in the drug resistance of ESBLs-EC, and this effect may be mediated by increasing PER-β-lactamases.

Keywords:

• GLO1
• bacterial drug resistance
• extended-spectrum β-lactamases
• Escherichia coli
• PER type

Ethical Statement

The study was conducted in accordance with the Declaration of Helsinki. Anonymized clinical samples obtained during routine hospital procedures were used for this study.

Acknowledgments

This study was supported by the Social Development Project of Jilin Science and Technology Department (No.20190303162SF) to Ke Wang, Advanced Program of Jilin Science and Technology Department (No.20191102012YY) to Ke Wang, Jilin Science and Technology Department (No.20200708083YY) to Ke Wang and Health Special Project of Jilin Provincial Finance Department to He Ma (No.2020SCZT070 and No.2020SCZT015). The funders had no role in the study design, data collection and analysis, interpretation of data, decision to publish, or writing the manuscript.

Author Contributions

All authors made significant contributions to conception, study design, execution, acquisition of data, or analysis and interpretation of data; took part in drafting the article or revising it critically for important intellectual content; gave final approval of the version to be published; have agreed on the journal submitted; and agree to be accountable for all aspects of the work.

Disclosure

All the authors have no competing interests to declare.