The kinetic properties of the carboxy terminal domain of the Bacillus licheniformis 749/I BlaR penicillin-receptor shed a new light on the derepression of beta-lactamase synthesis
[en] To study the properties of the BlaR penicillin-receptor involved in the induction of the Bacillus licheniformis beta-lactamase, the water-soluble carboxy terminal domain of the protein (BlaR-CTD) was overproduced in the periplasm of Escherichia coli JM105 and purified to protein homogeneity. Its interactions with various beta-lactam antibiotics were studied. The second-order acylation rate constants k(2)/K' ranged from 0.0017 to more than 1 muM(-1) s(-1) and the deacylation rate constants were lower than 4x10(-5) s(-1) . These values imply a rapid to very rapid formation of a stable acylated adduct. BlaR-CTD is thus one of the most sensitive penicillin-binding proteins presently described. In the light of these results, the kinetics of beta-lactamase induction in Bacillus licheniformis were re-examined. When starting with a rather high cell density, a good beta-lactamase substrate such as benzylpenicillin is too sensitive to beta-lactamase-mediated hydrolysis to allow full induction. By contrast, a poor beta-lactamase substrate (7-aminocephalosporanic acid) can fully derepress beta-lactamase expression under conditions where interference of the antibiotic with cell growth is observed. These results suggest that acylation of the penicillin receptor is a necessary, but not sufficient, condition for full induction.
Duval, Valérie; Université de Liège - ULiège > Centre d'Ingénierie des Protéines
Swinnen, Marc; Université de Liège - ULiège > Centre d'Ingénierie des Protéines
Lepage, Sophie; Université de Liège - ULiège > Centre d'Ingénierie des Protéines
Brans, Alain ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Granier, Benoit; Université de Liège - ULiège > Centre d'Ingénierie des Protéines
Franssen, Christine ; Université de Liège - ULiège > Centre d'Ingénierie des Protéines
Frère, Jean-Marie ; Université de Liège - ULiège > Centre d'Ingénierie des Protéines
Joris, Bernard ; Université de Liège - ULiège > Département des sciences de la vie > Physiologie et génétique bactériennes
Language :
English
Title :
The kinetic properties of the carboxy terminal domain of the Bacillus licheniformis 749/I BlaR penicillin-receptor shed a new light on the derepression of beta-lactamase synthesis
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