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Journal of Antimicrobial Chemotherapy (1988) 22, 613-621
© 1988 The British Society for Antimicrobial Chemotherapy

research-article

Activity of ß-lactamase in Pseudomonas aeruginosa as studied by HPLC

A. J. de Neeling*,a, A. Rutgersa, C. S. Schota and E. E. Stobberinghb

aLaboratory for Chemotherapy, National Institute of Public Health and Environmental Protection P.O. Box 1, 3720 BA Bilthoven, The Netherlands bDepartment of Medical Microbiology, University of Limburg P.O. Box 616. 6200 MD Maastricht, The Netherlands

Received 4 January 1988; returned 21 June 1988; accepted 21 June 1988

*Corresponding author

Mutants with increased resistance were selected from a clinical isolate of Pseudomonas aeruginosa using ceftazidime, piperadillin and carbenicillin. The MICs of these antibiotics and of ticarcillin were determined for the parent strain and for the selected mutants. Subsequently, cell-free extracts of the strains were prepared and the rates of hydrolysis of several ß-lactam substrates by the extracts were determined by HPLC procedures. It appeared possible to determine ß-lactamase activities in the crude cell extracts at the low substrate concentrations which may be attainable in the periplasm of Gram-negative bacteria. It is concluded that the increased drug MICs for mutants selected with ceftazidime or piperacillin, but not for those selected with carbenicillin, were caused by increased chromosomal ß-lactamase activity.


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