Hidekazu Suginaka

Penicillin-Resistant Mechanisms in Pseudomonas aeruginosa: Binding of Penicillin to Pseudomonas aeruginosa KM 338

A comparison of the binding of radioactive penicillin G to whole cells and the membrane fraction derived from Pseudomonas aeruginosa KM 338 was made. This organism has intrinsic resistance to penicillin. The binding to the membrane fraction which catalyzed peptidoglycan synthesis followed saturation type kinetics and saturation was achieved at approximately 2 nmol of penicillin G per ml, whereas binding to the whole cells was entirely of the nonsaturation type. The binding of carbenicillin to the membrane fraction was determined by competition between radioactive penicillin G and unlabeled carbenicillin for the binding sites. It was bound at the same sites in almost the same manner. When whole cells were pretreated with high concentration of unlabeled penicillin G or carbenicillin, the subsequent binding of radioactive penicillin G to the membrane fraction from carbenicillin-treated cells was entirely nonspecific, but with penicillin G-pretreated cells it was still specific. There was apparently specific binding of radioactive penicillin G to ethylenediaminetetraacetate-treated cells. P. aeruginosa KM 338 had an extremely low activity of β-lactamase compared with other enzyme-producing organisms. This enzyme from P. aeruginosa KM 338 was of the cephalosporinase type. These data indicate that penicillin resistance of P. aeruginosa KM 338 may be a consequence of the development of a permeability barrier which prevents the antibiotic from reaching its sites of action in the cytoplasmic membrane.

beta-Lactam resistance in Serratia marcescens: comparison of action of benzylpenicillin, Apalcillin, Cefazolin, and ceftizoxime.

The intrinsic mechanisms of resistance to beta-lactam antibiotics in Serratia marcescens IFO 12648 were investigated, comparing the action of benzylpenicillin, apalcillin, cefazolin, and ceftizoxime. The minimal inhibitory concentrations for this strain were 1,600, 3.13, 6,400, and 0.05 microgram/ml, respectively. The addition of ethylenediaminetetraacetic acid markedly reduced the minimal inhibitory concentrations of benzylpenicillin and cefazolin, whereas those of apalcillin and ceftizoxime were not influenced. S. marcescens IFO 12648 produced only a low level of beta-lactamase activity constitutively, and the production was considerably increased by the addition of benzylpenicillin. Cefazolin was hydrolyzed rapidly by beta-lactamase activity, whereas benzylpenicillin, apalcillin, and ceftizoxime were poorly hydrolyzed. Peptidoglycan synthesis in ether-treated strain IFO 12646 cells was inhibited by a concentration of ceftizoxime markedly lower than that of cefazolin and by a concentration of apalcillin moderately lower than that of benzylpenicillin.

Mechanism of Glucan‐Induced Agglutination in Streptococcus mutans*

The binding of radioactive glucan to Streptococcus mutatis cells, which are agglutinated by dextrans, was examined. The glucan was synthesized from sucrose by extracellular glucosyltransferases from S. mutans FA‐1 and was highly branched at C‐3 and C‐6 of D‐glucose residues, containing 17% of α(1→3)inter‐chain residues. Binding of glucan to whole cells of S. mutans OMZ‐176, which were agglutinated by addition of glucan or Dextran T2000, was irreversible and followed saturation type kinetics; saturation was achieved at approximately 110 ng of glucan per ml. About 14 ng of glucan were bound per mg of the cells at the saturated concentration. The heated cells of this organism, however, had a relatively low ability of glucan‐binding, compared with the freshly prepared and lyophilized cells. Binding to the heated cells was entirely of a non‐saturation type. Binding of Dextran T2000 or T10 was determined by competition between the labeled glucan and unlabeled Dextrans for the binding site(s). Both Dextrans and glucan from S. mutans FA‐1 were bound to the same site(s). Other organisms, which did not undergo glucan‐ and Dextran‐induced agglutination, had a relatively lower ability of glucan‐binding than S. mutans, which was agglutinated.

Effect of 6059-S, a novel oxacephem, on cross-linking reaction of peptidoglycan biosynthesis in Escherichia coli, Pseudomonas aeruginosa and Serratia marcescens

The effect of 6059-S, a novel 1-oxacephem, on peptidoglycan synthesis was investigated using ether-treated cells of Escherichia coli K 12, Pseudomonas aeruginosa KM 338 and Serratia marcescens IFO 12648. The cross-linking reaction of peptidoglycan synthesis in these organisms was inhibited by markedly low concentration of 6059-S.