Energy conservation in aerobically grown Staphylococcus aureus.

The present studies provide new data on the involvement of menaquinol oxidases in substrate oxidation and energy conservation in aerobically grown, resting cells of Staphylococcus aureus 17810R, starved of endogenous energy reserves and supplemented with glutamate or L-lactate. These cells were energetically competent, since they oxidized both substrates, generated an electrochemical proton gradient (deltamuH+) and synthesized ATP via oxidative phosphorylation. Studies with KCN showed that: (i) L-lactate oxidation occurred via two terminal menaquinol oxidases - the ba3-type sensitive to low KCN and the bo-type insensitive to cyanide, (ii) glutamate oxidation proceeded via the bo-type oxidase, and (iii) ATP synthesis with glutamate or L-lactate was coupled only to the bo-type oxidase. Also in glucose-grown cells oxidizing L-lactate, ATP synthesis was coupled to the highly repressed bo-type oxidase. It is suggested that in the respiratory chain of strain 17810R two energy coupling sites may be present: in the complex of NADH-menaquinone oxidoreductase and in the complex of the bo-type menaquinol oxidase. The rate of ATP synthesis was similar with both substrates, but the rate of their oxidation differed significantly: the P/O ratios were 1.5 and 0.03 with glutamate and L-lactate, respectively. CCCP accelerated glutamate oxidation by 50% but was without effect on L-lactate oxidation. In cell lysates, the rates of NADH and L-lactate oxidation were equal. It is concluded that in whole cells of S. aureus 17810R oxidation of NADH derived from glutamate breakdown is tightly coupled to phosphorylation, while L-lactate oxidation seems to be rather loosely coupled.

Effect of Cd2+ on phosphate uptake by cadmium-resistant and cadmium-sensitive Staphylococcus aureus.

The effect of Cd2+ on phosphate (Pi) uptake was investigated in the growing cells of Cd(2+)-resistant Staphylococcus aureus 1781OR and Cd(2+)-sensitive S. aureus 17810S. Inhibitor and ionophore studies showed that 32Pi uptake in the two strains occurred via the Pi porter down pH gradient (delta pH) generated by the respiratory chain. Cd2+ inhibited 32Pi uptake in the cadmium-sensitive strain 1781OS at all concentrations used (10 microM-1 mM). In strain 1781OR, possessing the plasmid-coded Cd2+ efflux system, 10-100 microM Cd2+ did not inhibit 32Pi uptake. Even at 1 mM Cd2+, inhibition of 32Pi uptake in strain 1781OR was reversed when the external Cd2+ was chelated with cysteine and activity of Cd2+ efflux system was restored. Cd2+ efflux induced by cysteine was energized either by membrane potential (delta psi) or by delta pH, which indicated that electrochemical gradient of protons (delta mu H+) was required for this efflux.

Plasmid-linked protection of [14C]-glutamate transport and its oxidation against Cd2+ in Staphylococcus aureus.

The effect of Cd2+ on [14C]-glutamate transport energized by endogenous respiration and on glutamate oxidation was studied in the Cd2+-resistant and -sensitive Staphylococcus aureus strains. The results indicate that these processes are protected against 10 microM Cd2+ in the Cd2+ resistant strain 17810R by the 2H+/Cd2+ antiporter encoded by the cadA determinant located on a penicillinase plasmid p II17810. Even at 100 microM Cd2+, glutamate oxidation was only partially inhibited in this organism and this inhibition appeared to be reversible. In the plasmidless variant strain 17810S, which lacks the 2H+/Cd2+ antiporter, both [14C]-glutamate transport and its oxidation was blocked by Cd2+ at 10 or 100 microM. In this strain, Cd2+-mediated inhibition of glutamate oxidation was irreversible. Energetics of glutamate transport in both strains was analyzed.