|
Author:
Date:
2015-09-05
[Abstract] Under aerobic conditions, Staphylococcus aureus (S. aureus) primarily metabolizes glucose to acetic acid. Although normally S. aureus is able to re-utilize acetate as a carbon source following glucose exhaustion, significantly high levels of acetate in the culture media may not only be growth inhibitory but also potentiates cell death in stationary phase cultures by a mechanism dependent on cytoplasmic acidification. One consequence of acetic acid toxicity is the production of reactive oxygen species (ROS). The present protocol describes the detection of ROS in S. aureus undergoing cell death by electron paramagnetic resonance (EPR) spectroscopy. Using 1-hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine (CMH) as a cell permeable spin probe, we ...
[摘要] 在有氧条件下，金黄色葡萄球菌（金黄色葡萄球菌）主要将葡萄糖代谢为乙酸。虽然通常是。金黄色葡萄球菌能够在葡萄糖耗竭之后再利用乙酸盐作为碳源，培养基中显着高水平的乙酸盐不仅可以是生长抑制性的，而且还可以通过依赖于细胞质的机制增强稳定期培养物中的细胞死亡酸化。乙酸毒性的一个后果是活性氧（ROS）的产生。本协议描述了ROS的检测。金黄色葡萄球菌通过电子顺磁共振（EPR）光谱法进行细胞死亡。使用1-羟基-3-甲氧基羰基-2,2,5,5-四甲基吡咯烷（CMH）作为细胞可渗透的自旋探针，我们证明检测由细菌产生的各种氧自由基。虽然标准化了 s。 aureus ，这里描述的方法应该很容易适应其他细菌物种。该协议改编自托马斯（Thomas）等人（2014）和托马斯（Thomas）等人（2010）。
|