Purification and properties of a constitutive beta-lactamase from Pseudomonas aeruginosa strain Dalgleish.
1. The beta-lactamase (penicillin amido-beta-lactamhydrolase EC 3.5.2.6) appeared to be periplasmic rather than truly intracellular, since it was released by freeze-thawing without gross morphological changes in the cell. 2. The partially purified enzyme had pI between 5.0 and 5.5, mol. wt 32 000 and a broad pH vs activity profile with a maximum at pH 8. 3. The cephalosporins tested were hydrolysed less rapidly than most of the penicillins, and the Km values for penicillins were lower than for cephalosporins. However cloxacillin was hydrolysed very slowly although it was strongly bound. The substrate-induced inactivation common to many beta-lactamases was particularly marked with cephaloridine and cloxacillinmthe cloxacillin-induced inactivation was shown to be reversible.
D002509:Cephaloridine / Q000494:pharmacology; D002511:Cephalosporins; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003023:Cloxacillin / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005615:Freezing; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007700:Kinetics; D008970:Molecular Weight; D010405:Penicillinase / Q000302:isolation & purification / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D013329:Structure-Activity Relationship; D013347:Subcellular Fractions / Q000201:enzymology
D002511:Cephalosporins; D010405:Penicillinase; D002509:Cephaloridine; D003023:Cloxacillin