Changes in the Activity of Antioxidant Systems of Escherichia coli under Phosphate Starvation

Abstract—Changes in the activity of antioxidant systems in Escherichia coli during phosphate starvation have been studied. It is shown that starvation was accompanied by a decrease in the intensity of respiration, an increase in the rate of superoxide production, and a decrease in the level of ATP. Simultaneously, there was a decrease in H₂O₂ in the medium and a significant increase in the expression of the katG and katE genes which encode the HPI and HPII catalases, respectively. At the same time, there was no drop in the membrane potential, which may indicate the retention of normal membrane activity in starving cells. It has been shown for the first time that the transition of E. coli to phosphate starvation is accompanied by significant changes in the status of glutathione. The most important of these are associated with a decrease in the level of reduced glutathione in the medium (GSH_out) and with a simultaneous increase in its content in the cytoplasm (GSH_in), as well as a shift in the GSH_in to oxidized glutathione form (GSSG_in) ratio towards reductive values, and GSH_out/GSSG_out towards oxidative values. Among the mutants used in the work, the gor trxB double mutant, which is deficient in the synthesis of glutathione reductase and thioredoxin reductase, showed the most pronounced distinctive features. Compared to the parental strain, this mutant showed a multiple higher expression of katG::lacZ, the highest level of oxidized intra- and extracellular glutathione, and, accordingly, the lowest GSH/GSSG ratio in both compartments. In general, the data we obtained indicate that during phosphate starvation the interaction of the glutathione redox-system and regulons that control protection against reactive oxygen species creates conditions that allow maintaining the concentration of ROS below the toxic level. As a result, phosphate-starved E. coli cells can maintain high viability for a long period of time, which allows them to quickly resume growth after the addition of phosphate.