Study of linkage between glutathione pathway and the antibiotic resistance of Escherichia coli from patients' swabs

谷胱甘肽途径与患者拭子大肠杆菌抗生素耐药性的关联研究

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作者:Marketa Kominkova, Petr Michalek, Kristyna Cihalova, Roman Guran, Natalia Cernei, Lukas Nejdl, Kristyna Smerkova, Simona Dostalova, Dagmar Chudobova, Zbynek Heger, Radek Vesely, Jaromir Gumulec, Jindrich Kynicky, Kledi Xhaxhiu, Ondrej Zitka, Vojtech Adam, Rene Kizek1

Abstract

In this work, we focused on the differences between bacterial cultures of E. coli obtained from swabs of infectious wounds of patients compared to laboratory E. coli. In addition, blocking of the protein responsible for the synthesis of glutathione (γ-glutamylcysteine synthase-GCL) using 10 mM buthionine sulfoximine was investigated. Each E. coli showed significant differences in resistance to antibiotics. According to the determined resistance, E. coli were divided into experimental groups based on a statistical evaluation of their properties as more resistant and more sensitive. These groups were also used for finding the differences in a dependence of the glutathione pathway on resistance to antibiotics. More sensitive E. coli showed the same kinetics of glutathione synthesis while blocking GCL (Km 0.1 µM), as compared to non-blocking. In addition, the most frequent mutations in genes of glutathione synthetase, glutathione peroxidase and glutathione reductase were observed in this group compared to laboratory E.coli. The group of "more resistant" E. coli exhibited differences in Km between 0.3 and 0.8 µM. The number of mutations compared to the laboratory E. coli was substantially lower compared to the other group.

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