Max-Planck-Institute for Dynamics of Complex Technical Systems, Sandtorstraße, Magdeburg, Germany.
PLoS One. 2018 Apr 3;13(4):e0194699. doi: 10.1371/journal.pone.0194699. eCollection 2018.
The electron transport chain of E. coli contains three different quinone species, ubiquinone (UQ), menaquinone (MK) and demethylmenaquinone (DMK). The content and ratio of the different quinone species vary depending on the external conditions. To study the function of the different quinone species in more detail, strains with deletions preventing UQ synthesis, as well as MK and/or DMK synthesis were cultured under aerobic and anaerobic conditions. The strains were characterized with respect to growth and product synthesis. As quinones are also involved in the control of ArcB/A activity, we analyzed the phosphorylation state of the response regulator as well as the expression of selected genes.The data show reduced aerobic growth coupled to lactate production in the mutants defective in ubiquinone synthesis. This confirms the current assumption that ubiquinone is the main quinone under aerobic growth conditions. In the UQ mutant strains the amount of MK and DMK is significantly elevated. The strain synthesizing only DMK is less affected in growth than the strain synthesizing MK as well as DMK. An inhibitory effect of MK on aerobic growth due to increased oxidative stress is postulated.Under fermentative growth conditions the mutant synthesizing only UQ is severely impaired in growth. Obviously, UQ is not able to replace MK and DMK during anaerobic growth. Mutations affecting quinone synthesis have an impact on ArcA phosphorylation only under anaerobic conditions. ArcA phosphorylation is reduced in strains synthesizing only MK or MK plus DMK.
大肠杆菌的电子传递链包含三种不同的醌类物质,泛醌(UQ)、甲萘醌(MK)和去甲甲萘醌(DMK)。不同醌类物质的含量和比例会根据外部条件而变化。为了更详细地研究不同醌类物质的功能,我们培养了不能合成 UQ 以及(或)不能合成 MK 和/或 DMK 的缺失菌株,在需氧和厌氧条件下进行培养。我们对这些菌株的生长和产物合成情况进行了特征描述。由于醌类物质也参与 ArcB/A 活性的调控,我们分析了反应调节蛋白的磷酸化状态以及选定基因的表达情况。数据显示,在不能合成泛醌的突变体中,有氧生长受到抑制,伴随着乳酸的产生。这证实了当前的假设,即在有氧生长条件下,泛醌是主要的醌类物质。在 UQ 突变株中,MK 和 DMK 的含量显著增加。只合成 DMK 的菌株在生长方面的受影响程度比同时合成 MK 和 DMK 的菌株要小。由于氧化应激增加,MK 对有氧生长具有抑制作用。在发酵生长条件下,只合成 UQ 的突变株在生长方面受到严重损害。显然,在厌氧生长过程中,UQ 无法替代 MK 和 DMK。影响醌类物质合成的突变只有在厌氧条件下才会对 ArcA 磷酸化产生影响。只合成 MK 或 MK 和 DMK 的菌株中,ArcA 的磷酸化减少。