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Induction of Escherichia coli hydroperoxidase I by acetate and other weak acids.

机译:醋酸盐和其他弱酸诱导大肠杆菌氢过氧化物酶I。

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摘要

Escherichia coli produces two independently regulated hydroperoxidases (catalases) that protect the cell from toxic concentrations of hydrogen peroxide. Hydroperoxidase I (HPI) is induced by hydrogen peroxide in an OxyR-dependent manner, while hydroperoxidase II (HPII) synthesis is regulated by an alternative sigma factor called RpoS (KatF). The activities of both hydroperoxidases increase as exponentially growing cells enter stationary phase. In this study, we examined the growth phase-dependent expression of HPI. Treatment of early-exponential-phase cells with spent culture supernatant resulted in induction of HPI synthesis. Extracellular levels of hydrogen peroxide, accumulating in the culture supernatant during late exponential phase, were found to be lower than the concentrations normally required to induce OxyR-dependent synthesis of HPI. This finding suggested that factors other than hydrogen peroxide may play a role in HPI expression. Weak acids such as acetate, which accumulate in culture supernatant and have been implicated in the regulation of HPII, caused a sixfold increase in HPI expression. Increases in HPI synthesis, mediated by weak acids and spent culture fluid supernatant, could be prevented by chloramphenicol, indicating that de novo protein synthesis is required for induction. Expression studies using a plasmid-borne lacZ transcriptional fusion to katG, the structural gene for HPI, indicated that growth phase-dependent regulation of HPI occurs primarily at the level of transcription and is dependent on RpoS. These results suggest that there may be a common regulatory mechanism of HPI and HPII expression in addition to previously described independent control mechanisms.
机译:大肠杆菌产生两种独立调节的氢过氧化物酶(催化酶),可保护细胞免受毒性浓度的过氧化氢的伤害。氢过氧化物酶I(HPI)由过氧化氢以OxyR依赖性方式诱导,而氢过氧化物酶II(HPII)的合成则受称为RpoS(KatF)的另类sigma因子调节。随着指数增长的细胞进入固定期,两种氢过氧化物酶的活性都增加。在这项研究中,我们检查了HPI的生长阶段依赖性表达。用用过的培养上清液处理早期指数期细胞导致诱导HPI合成。发现在指数末期在培养上清液中积累的细胞外过氧化氢水平低于诱导OxyR依赖性HPI合成所需的正常浓度。该发现表明,过氧化氢以外的因素可能在HPI表达中起作用。弱酸(例如乙酸盐)会在培养上清液中积累,并与HPII的调节有关,从而导致HPI表达增加六倍。氯霉素可以防止由弱酸和废培养液上清液介导的HPI合成增加,这表明诱导需要从头合成蛋白质。使用质粒载体lacZ转录融合蛋白对HPI的结构基因katG进行的表达研究表明,HPI的生长阶段依赖性调节主要发生在转录水平,并且依赖于RpoS。这些结果表明,除了先前描述的独立控制机制外,可能还有HPI和HPII表达的共同调控机制。

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