Julian William T, Vasilchenko Anastasia V, Shpindyuk Daniil D, Poshvina Darya V, Vasilchenko Alexey S
Laboratory of Antimicrobial Resistance, Institute of Environmental and Agricultural Biology (X-BIO), Tyumen State University, 625003 Tyumen, Russia.
All-Russian Institute of Plant Protection, 196608 St. Petersburg, Russia.
Biomolecules. 2020 Dec 25;11(1):13. doi: 10.3390/biom11010013.
2,4-Diacetylphloroglucinol (2,4-DAPG) is a well-known bacterial secondary metabolite, however, its mechanism of inhibitory and subinhibitory action on bacterial cells is still poorly understood. The mechanism of 2,4-DAPG action on model bacterial strains was investigated using fluorescent spectroscopy and the action of the antibiotic was found to involve a rapid increase in membrane permeability that was accompanied by a reduction in its viability in nutrient-poor medium. At the same time, antibacterial action in nutrient-rich medium developed for several hours. Atomic force microscopy demonstrated time-dependent disturbances in the outer membrane of when exposed to 2,4-DAPG, while cells have been visualized with signs of intracellular leakage. In addition, 2,4-DAPG inhibited the metabolic activity of and bacterial cells in mature biofilms. Observed differences in the antibiofilm activity were dependent upon antibiotic concentration. The intracellular targets of the action of 2,4-DAPG were assessed using bacterial biosensors with inducible bioluminescence corresponding to DNA and protein damage. It was unable to register any positive response from either sensor. As a result, the bactericidal action of 2,4-DAPG is believed to be associated with the destruction of the bacterial barrier structures. The subinhibitory effect of 2,4-diacetylphloroglucinol was tested on quorum-sensing mediated processes in . Subinhibitory concentrations of 2,4-DAPG were found to lower the biosynthesis of acyl-homoserine lactones in in a dose-dependent manner. Further investigation elucidated that 2,4-DAPG inhibits the metabolic activity of bacteria without affecting their viability.
2,4-二乙酰基间苯三酚(2,4-DAPG)是一种著名的细菌次级代谢产物,然而,其对细菌细胞的抑制和亚抑制作用机制仍知之甚少。利用荧光光谱法研究了2,4-DAPG对模式细菌菌株的作用机制,发现该抗生素的作用涉及膜通透性的快速增加,同时在营养贫乏培养基中其活力降低。与此同时,在营养丰富的培养基中抗菌作用会持续数小时。原子力显微镜显示,当暴露于2,4-DAPG时,细菌外膜会出现时间依赖性干扰,同时观察到细胞有细胞内泄漏的迹象。此外,2,4-DAPG抑制成熟生物膜中细菌和细菌细胞的代谢活性。观察到的抗生物膜活性差异取决于抗生素浓度。使用对应于DNA和蛋白质损伤的可诱导生物发光的细菌生物传感器评估了2,4-DAPG作用的细胞内靶点。两种传感器均未检测到任何阳性反应。因此,认为2,4-DAPG的杀菌作用与细菌屏障结构的破坏有关。测试了2,4-二乙酰基间苯三酚对群体感应介导过程的亚抑制作用。发现2,4-DAPG的亚抑制浓度以剂量依赖的方式降低了细菌中酰基高丝氨酸内酯的生物合成。进一步研究表明,2,4-DAPG抑制细菌的代谢活性而不影响其活力。