Institute of Ecology and Genetics of Microorganisms, Perm Federal Research Center, Russian Academy of Sciences, Golev Street 13, 614081, Perm, Russia.
Res Microbiol. 2022 Jul-Sep;173(6-7):103954. doi: 10.1016/j.resmic.2022.103954. Epub 2022 May 11.
In most previous studies the sensitivity of Escherichia coli outer membrane mutants to ciprofloxacin (CF) was studied by MIC method. In the present work, the early response of these mutants to CF was studied using physiological and biochemical methods and electrochemical sensors. The use of sensors made it possible to monitor dissolved oxygen, potassium and extracellular sulfide continuously directly in growing cultures in real time. In the absence of CF, no significant differences were found between the mutants deficient in porin OmpF and lipopolysaccharide (LPS) and the parent. The only exception was 5-6 times higher extracellular glutathione and 1.5-3 times lower intracellular glutathione in the lpcA compared to the parent and the ompF. Ciprofloxacin inhibited growth, respiration, membrane potential and K consumption, which was less pronounced in both mutants compared to the parent. Changes in these parameters correlated with each other, but not with survival. A reversible increase in sulfide level was observed at 3 μg ml CF in the parent, at 20 μg ml CF in ompF and was absent in lpcA at all concentrations. The data obtained show that the use of electrochemical sensors can provide a more complete understanding of the early response of bacteria to CF.
在大多数先前的研究中,通过 MIC 法研究了大肠埃希氏菌外膜突变体对环丙沙星 (CF) 的敏感性。在本工作中,使用生理生化方法和电化学传感器研究了这些突变体对 CF 的早期反应。使用传感器使得可以实时直接在生长培养物中连续监测溶解氧、钾和细胞外的硫化物。在没有 CF 的情况下,缺失孔蛋白 OmpF 和脂多糖 (LPS) 的突变体与亲本之间没有发现显著差异。唯一的例外是与亲本和 ompF 相比,lpcA 中外源谷胱甘肽高 5-6 倍,细胞内谷胱甘肽低 1.5-3 倍。环丙沙星抑制生长、呼吸、膜电位和 K 消耗,与亲本相比,这两种突变体的抑制作用较小。这些参数的变化相互关联,但与存活率无关。在亲本中,在 3μg/ml CF 时观察到硫化物水平可逆增加,在 ompF 中在 20μg/ml CF 时观察到,而在 lpcA 中在所有浓度下均不存在。所得数据表明,电化学传感器的使用可以更全面地了解细菌对 CF 的早期反应。