R-Mediated β-Lactamases and Episomal Resistance to the β-Lactam Drugs in Different Bacterial Hosts

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Abstract
Ten penicillinase plasmids of varying taxonomic origin were studied after transfer to a variety of bacterial hosts. Nine of the ten plasmids specified enzymes with the following identical, or very similar, properties: substrate profile, molecular weight, susceptibility to heat and inhibitors, and electrophoretic mobility, i.e., TEM-like enzymes. The tenth R-mediated β-lactamase was a cephalosporinase. Plasmids with TEM-like enzymes mediated resistance patterns identical towards the β-lactam drugs, whereas the resistance pattern of the cephalosporinase plasmid was distinctly different. Expression of enzyme and resistance had a dual R-factor and host specificity. Escherichia coli K-12 and Salmonella typhi constituted one group of the same R-factor phenotype expressions. Most, but not all, penicillinase plasmids exhibited in Proteus PM1 a considerably lower order of β-lactamase activity and an even lower order of resistance to the β-lactam drugs than the previous two hosts. This difference was most pronounced for the resistance to carbenicillin, which was mediated by the plasmids specifying the synthesis of TEM-like enzymes. Release by osmotic shock was complete in the host E. coli K-12 for the TEM-like enzymes, but was lower for the cephalosporinase and minimal or negative in the PM1 host. Crypticity factor for benzylpenicillin, ampicillin, and carbenicillin was not related to the increase in resistance mediated by the penicillinase plasmids in both K-12 and PM1 hosts. Inoculum size effects for the penicillins and 6-aminopenicillanic acid were higher in PM1 than in K-12 R + cultures. The expression of penicillinase plasmids in wild-type bacteria was strain specific and not species specific. For two plasmids of different phenotypes for β-lactamase activity (and resistance) in K-12 and PM1 hosts, a positive correlation was found between their phenotype and the relative amount of episomal deoxyribonucleic acid, as detected by ethidium bromide density gradient centrifugation. This is interpreted as indicating differences in the mode of replication of the plasmids in the two hosts.