Kinetics and significance of the activity of the Sabath and Abrahams' beta-lactamase of Pseudomonas aeruginosa against cefotaxime and cefsulodin.
All Pseudomonas aeruginosa strains produce Sabath and Abrahams' (SA) enzyme, as inducible beta-lactamase. Hydrolysis of cefotaxime by this enzyme, although slow in terms of Vmax was efficient when the parameter Vmax/Km (physiological efficiency (Pollock, 1965)) was considered. Hydrolysis of cefsulodin was not detectable in assays used and enzyme binding (Ki) was poor, ensuring a very low physiological efficiency. Physiological efficiency represents a measure of enzyme function under low substrate (antibiotic) conditions, as apply in the periplasm; consequently SA enzyme might protect the cell against cefotaxime, but not cefsulodin. This depends on enzyme induction and retention within the periplasm. Agar checkerboard studies indicated the SA inducer cephaloridine antagonized the activity of cefotaxime against most (12/14) Ps. aeruginosa strains but cefsulodin against only a minority (2/14). Cephaloridine/cefotaxime antagonism was lost in uninducible (SAI-) or constitutive (SAIcon) mutants where SA expression was independent of cephaloridine concentration. This indicated the antagonism was SA dependent. Cefotaxime plate MICs against parent SAI+ and uninducible SAI- organisms were similar indicating cefotaxime did not induce SA enzyme in these tests. Cefotaxime was however much less active against the SAI+ organism than the SAI- when log phase broth cultures were exposed to antibiotic and incubation was extended to 30 h. This correlated with observed SA induction in the SAI+ organism. Cefsulodin MICs against SAI+ and SAI- organisms were similar and no difference existed in activity, over long periods against broth cultures. Overall, results indicated that SA enzyme, if induced, constituted a defense against cefotaxime but not cefsulodin and this correlated with the physiological efficiency results. To what degree induction occurs in vivo during cefotaxime therapy of pseudomonal infections remains unknown.
D002439:Cefotaxime / Q000378:metabolism*; D002441:Cefsulodin; D002509:Cephaloridine / Q000378:metabolism; D002510:Cephalosporinase / Q000378:metabolism*; D002511:Cephalosporins / Q000378:metabolism*; D004352:Drug Resistance, Microbial; D004790:Enzyme Induction; D007700:Kinetics; D009154:Mutation; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D001618:beta-Lactamases / Q000378:metabolism*
D002511:Cephalosporins; D002510:Cephalosporinase; D001618:beta-Lactamases; D002509:Cephaloridine; D002439:Cefotaxime; D002441:Cefsulodin