Damaging effects of ethylenediaminetetra-acetate and penicillins on permeability barriers in Gram-negative bacteria

1. The permeability barrier against benzylpenicillin has been found to be passive in four strains of penicillinase-producing Gram-negative bacteria (three of Klebsiella aerogenes and one of Escherichia coli). 2. If the three K. aerogenes strains are grown in the presence of sub-inhibitory concentrations of benzylpenicillin, ampicillin or phenethicillin the resultant bacterial cells have deficient permeability barriers. Concentrations of ampicillin or benzylpenicillin less than one-tenth of those required to inhibit growth cause destruction of more than half the permeability barrier in these strains. 3. Benzylpenicillin, ampicillin and phenethicillin have no effect upon the permeability barriers of resting cells from the three K. aerogenes strains. 4. Treatment of resting cells with trisodium EDTA, although failing to sensitize K. aerogenes to lysozyme, severely damages permeability barriers in this species. 5. The magnesium and calcium salts of EDTA do not have the same capacity as the sodium salt for causing damage to permeability barriers in K. aerogenes and E. coli. Damage caused by trisodium EDTA can be at least partially reversed by treatment with Ca(2+) or Mg(2+) ions. It is suggested that EDTA damage is caused by removal of either Ca(2+) or Mg(2+) ions, or both, from the bacterial cell envelope. 6. Bacterial cells with deficient permeability barriers as a result of either growth in the presence of a penicillin or treatment with EDTA remain viable, and revert to their usual permeability after growth in nutrient broth.