Purification and properties of beta-lactamase from Proteus morganii.
The cephalosporin beta-lactamase was purified from a strain of Proteus morganii that showed resistance to beta-lactam antibiotics and produced the enzyme constitutively. The purified enzyme preparation gave a single protein band on polyacrylamide gel electrophoresis and consisted of a single polypeptide of molecular weight 38,000 to 40,000 from gel filtration of Sephadex G-100 and sodium dodecyl sulfate-acrylamide gel electrophoresis, its isoelectric point being pH 7.2 No cysteine residue was found in its amino acid composition. The specific activity was 190 mumol/min per mg of the purified enzyme protein for the hydrolysis of cephaloridine, the optimal pH was about 8.5 and the optimal temperature was 50 degrees C. Antibodies against the purified beta-lactamase inhibited not only the enzyme activity of the purified preparation, but also the enzyme activity of all of the other strains of P. morganii so far tested, regardless of whether the modes of their production were inducible or constitutive. None of the beta-lactamases produced by beta-lactam antibiotic-resistant strains of other species of Proteus was affected at all by the antibodies, thus showing that the purified cephalosporin beta-lactamase was of the species-specific type. The enzymological properties of the preparation have been compared with those of beta-lactamases derived from other gram-negative enteric bacteria.
D000581:Amidohydrolases / Q000302:isolation & purification; D000596:Amino Acids / Q000032:analysis; D002510:Cephalosporinase / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism; D002511:Cephalosporins / Q000378:metabolism / Q000494:pharmacology; D000939:Epitopes; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008970:Molecular Weight; D010403:Penicillin Resistance; D010406:Penicillins / Q000378:metabolism / Q000494:pharmacology; D010455:Peptides / Q000032:analysis; D011511:Proteus / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology; D013696:Temperature
D000596:Amino Acids; D002511:Cephalosporins; D000939:Epitopes; D010406:Penicillins; D010455:Peptides; D000581:Amidohydrolases; D002510:Cephalosporinase