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海洋来源的链霉菌次级代谢产物通过膜通透性对细菌分离物作用的机制。

Mechanism of action of secondary metabolites from marine-derived Streptoymces on bacterial isolates by membrane permeability.

机构信息

Applied Microbial Processes & Environmental Health Research Group (AMPEHREG), Faculty of Life Sciences, University of Benin, Private Mail Bag 1154, Benin City, 300283, Nigeria.

Applied Microbial Processes & Environmental Health Research Group (AMPEHREG), Faculty of Life Sciences, University of Benin, Private Mail Bag 1154, Benin City, 300283, Nigeria; Department of Microbiology, College of Natural and Applied Sciences, Western Delta University, Private Mail Bag 10 Oghara, 331101, Nigeria.

出版信息

Microb Pathog. 2020 Dec;149:104532. doi: 10.1016/j.micpath.2020.104532. Epub 2020 Sep 28.

Abstract

This study was designed to further investigate the potential mechanism of action of our previously characterized and identified marine-derived Streptomyces extracts (ESC003 and ESC012) on selected bacterial isolates from our culture bank. Time-kill kinetics, protein and lipid leakages assay, cell membrane permeability, phosphate and potassium ions efflux assay, extracellular adenosine triphosphate (ATP) concentration and membrane potential (MP) were all carried out using the marine-derived Streptomyces extract to determine and understand the probable mode of action at which the extract inhibit or kill bacterial cell. The MIC of ESC003 and ESC012 ranged from 0.16 to 6.25 mg/mL while the MBC ranged from 1.25 to >10 mg/mL. On the time-kill kinetics, a reduction in mean viable cell amount was detected at respective time studied. For the impermeability of the bacterial isolates, the relative electric conductivity increased with increase in concentration and time interval of exposure. As regards the protein leakage, lipid leakage, 260 nm absorbing materials leakage, phosphate and potassium ions efflux; considerable amount of these products were leaked with increase in concentration and time of exposure to the bacterial isolates. The extracellular ATP concentration from respective bacterial isolates increased appreciably with increased concentration of exposure with a simultaneous decrease in membrane potential. Findings from this study revealed that the Streptomyces extracts revealed a significant breakthrough against susceptible bacterial isolates via the permeability of the bacterial cell membrane, and thus resulted in the outflow of ATP, electrolytes, DNA materials, and proteins. These changes lead to disruption, and eventually cell death, which were proportional to a concurrent decrease in the viability of the bacterial cell.

摘要

本研究旨在进一步探讨我们之前从培养库中分离出的海洋衍生链霉菌提取物(ESC003 和 ESC012)对选定细菌分离物的潜在作用机制。采用海洋衍生链霉菌提取物进行时间杀伤动力学、蛋白质和脂质渗漏测定、细胞膜通透性、磷酸盐和钾离子外排测定、细胞外三磷酸腺苷(ATP)浓度和膜电位(MP)测定,以确定和理解提取物抑制或杀死细菌细胞的可能作用模式。ESC003 和 ESC012 的 MIC 范围为 0.16 至 6.25mg/mL,MBC 范围为 1.25 至>10mg/mL。在时间杀伤动力学中,在研究的相应时间检测到存活细胞数量减少。对于细菌分离物的通透性,相对电导率随浓度和暴露时间间隔的增加而增加。关于蛋白质渗漏、脂质渗漏、260nm 吸收材料渗漏、磷酸盐和钾离子外排;随着暴露于细菌分离物的浓度和时间的增加,这些产物的数量大量泄漏。来自各个细菌分离物的细胞外 ATP 浓度随着暴露浓度的增加而显著增加,同时膜电位降低。这项研究的结果表明,链霉菌提取物通过细菌细胞膜的通透性对敏感细菌分离物显示出显著的突破,从而导致 ATP、电解质、DNA 物质和蛋白质的外流。这些变化导致破坏,最终导致细胞死亡,这与细菌细胞活力的同步下降成正比。

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