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硫化物在大肠杆菌培养物中应激诱导 Eh 变化中的作用。

The role of sulfides in stress-induced changes of Eh in Escherichia coli cultures.

机构信息

Institute of Ecology and Genetics of Microorganisms, Russian Academy of Sciences, Golev street 13, 614081 Perm, Russia.

Institute of Ecology and Genetics of Microorganisms, Russian Academy of Sciences, Golev street 13, 614081 Perm, Russia.

出版信息

Bioelectrochemistry. 2018 Jun;121:11-17. doi: 10.1016/j.bioelechem.2017.12.012. Epub 2018 Jan 3.

Abstract

Real-time monitoring of the state of bacterial cultures is important in both experiment and biotechnology. Using Eh and sulfide sensors, we demonstrated that the abrupt reversible reduction in Eh (Eh jump), occurring during transition of E. coli from exponential growth to starvation and antibiotic-induced stresses, is the result of sulfide excretion from the cells. Changes in the potential of sensors had a two-phase mode. The potential reduced within 10-15min and returned within 10-30min. In the parental strain, maximum amplitudes of Eh jumps (ΔEh) were 25±2mV, 57±6mV and 36±7mV under isoleucine starvation, glucose depletion and ciprofloxacin exposure that corresponded to 43±3nM, 96±5nM and 140±1nM of sulfide, respectively. In the glutathione-deficient mutant (ΔgshA), ΔEh values and sulfide concentration increased 1.5-4 times compared to the parent. Stress-induced sulfide excretion occurred in the background of inhibition of growth and respiration and a decrease in the membrane potential. The formation of sulfide caused by cysteine desulfurization may be related with maintaining of cysteine homeostasis under conditions of slow metabolism. There was a close relationship between transmembrane fluxes of sulfide, cysteine and glutathione.

摘要

实时监测细菌培养物的状态在实验和生物技术中都很重要。我们使用 Eh 和硫化物传感器证明,当大肠杆菌从指数生长过渡到饥饿和抗生素诱导的应激时,Eh(Eh 跃变)的突然可逆降低是细胞排出硫化物的结果。传感器电位的变化呈两相模式。电位在 10-15 分钟内降低,并在 10-30 分钟内恢复。在亲本菌株中,在亮氨酸饥饿、葡萄糖耗尽和环丙沙星暴露下,Eh 跃变(ΔEh)的最大幅度分别为 25±2mV、57±6mV 和 36±7mV,分别对应于 43±3nM、96±5nM 和 140±1nM 的硫化物。在谷胱甘肽缺陷突变体(ΔgshA)中,与亲本相比,ΔEh 值和硫化物浓度增加了 1.5-4 倍。应激诱导的硫化物排泄发生在生长和呼吸抑制以及膜电位下降的背景下。半胱氨酸脱硫引起的硫化物形成可能与在代谢缓慢的条件下维持半胱氨酸的平衡有关。硫化物、半胱氨酸和谷胱甘肽的跨膜通量之间存在密切关系。

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