The stringent response gene rsh plays multiple roles in Novosphingobium pentaromativorans US6-1's accommodation to different environmental pollutants: Phenanthrene, copper and nZVI.

The rsh based stringent response system is widely employed by bacteria to cope with environmental stresses. However, how does the stringent response involve in bacterial accommodation to environmental pollutant is largely unexplored. In this study, to comprehensively understand the roles of rsh in Novosphingobium pentaromativorans US6-1's metabolism and accommodation to different pollutants, three distinct pollutants, phenanthrene, copper and nanoparticulated zero valent iron (nZVI) were selected as exposure substances. Results indicated that rsh played important roles in US6-1's multiplication and metabolism, including survival rate at stationary phase, amino acid and nucleotide metabolism, extracellular polymeric substance (EPS) production, redox homeostasis, etc. The deletion of rsh affected phenanthrene removal rates through regulating the multiplication of US6-1 and increasing the expression of degradation related genes. The rsh mutant showed higher resistance to copper than the wild type, largely due to higher EPS production and enhanced expression of copper resistance related genes. Finally, the rsh based stringent response helped maintain the redox homeostasis when US6-1 confronted nZVI particles that exerted oxidative stress, thereby improving the survival rate. Overall, this study provides firsthand data that rsh plays multiple roles in US6-1's accommodation to environmental pollutants. The stringent response system could be a powerful tool for environmental scientists and engineers to harness bacterial activities for bioremediation purposes.

Oxygen availability affects the synthesis of quorum sensing signal in the facultative anaerobe Novosphingobium pentaromativorans US6-1.

Bacterial populations rely on quorum sensing (QS) to coordinate their behaviors and are often challenged by the fluctuation in oxygen concentrations in their habitats. Oxygen is a crucial factor that affects bacterial metabolism in multiple ways. However, little is known about whether and how oxygen availability affects QS activities. To fill this gap, we used the facultative anaerobe Novosphingobium pentaromativorans US6-1 as a model system, and observed that the QS signal acyl homoserine-lactones (AHLs) were produced only in anoxic environments, such as biofilm, or liquid medium that initially contained less than 2 mg/L dissolved oxygen, but not in highly oxic environments. Comparative transcriptome analysis revealed that oxygen availability significantly affected the physiological activities in US6-1, including fatty acid metabolism, oxidative phosphorylation, citrate cycle, QS activities, and flagellar assembly. The absence of AHLs in the oxic culture was not due to degradation, but to the very low expression of the AHL synthase gene novI. High concentration of NADH during the middle log phase under static cultivation may be a trigger for AHL synthesis. This is the first report that production of AHLs is coupled with anoxic metabolism in a facultative anaerobe, which extends our knowledge on factors affecting bacterial QS occurrence. KEY POINTS: • AHL production is anoxic cultivation related. • Oxygen availability affects AHL synthesis by influencing novI expression. • Oxygen availability changes many metabolism activities including NADH production.