The PhoPR Two-Component System Responds to Oxygen Deficiency and Regulates the Pathways for Energy Supply in Corynebacterium Glutamicum

The PhoPR two-component system, a highly conserved system in corynebacteria and mycobacteria, is involved in the cellular response to environmental stress. When analysing the transcriptomic data of Corynebacterium glutamicum strains under different dissolved oxygen (DO) levels, PhoPR was found to be the most responsive two-component system to DO changes. Here, we systematically investigated the expression of PhoPR in response to different DO levels and its impact on genes related to global regulation and energy metabolism. Using Green fluorescent protein as a reporter, we confirmed that PhoPR was significantly upregulated upon decrease of DO. Through real-time quantitative PCR and electrophoretic mobility shift assay, we found that the effector protein PhoP directly activated glxR (encoding a global regulator), pfk and gapA (encoding the glycolytic enzymes) and ctaD (encoding cytochrome c in the electron transport chain), while downregulated aceE and gltA (encoding the TCA cycle enzymes). Overexpression of phoP or phoR resulted in a decreased intracellular NAD+/NADH ratio and increased intracellular ATP level, consistent with the gene expression changes regulated by PhoP. These results reveal the PhoPR system respond to oxygen deficiency and is responsible for the regulation of pathways involved in the sustainability of the energy levels required under low oxygen conditions. Our findings in this study not only provide new insights into the adaptation pathways of C. glutamicum in response to low oxygen conditions but also identify new possible genetic targets for the construction of the new cell factories aimed toward industrial applications.