Article (Scientific journals)
Method for estimation of low outer membrane permeability to beta-lactam antibiotics
Lakaye, Bernard; Dubus, Alice; Joris, Bernard et al.
2002In Antimicrobial Agents and Chemotherapy, 46 (9), p. 2901-2907
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Abstract :
[en] The outer membrane of gram-negative bacteria plays a major role in beta-lactam resistance as it slows down antibiotic entry into the periplasm and therefore acts in synergy with beta-lactamases and efflux systems. Up to now, the quantitative estimation of low outer membrane permeability by the method of Zimmermann and Rosselet was difficult because of the secreted and cell surface-associated beta-lactamases. The method presented here uses the acylation of a highly sensitive periplasmic penicillin-binding protein (PBP) (BlaR-CTD) to assess the rate of beta-lactam penetration into the periplasm. The method is dedicated to measurement of low permeability and is only valid when the diffusion rate through the outer membrane is rate limiting. Cytoplasmic membrane associated PBPs do not interfere since they are acylated after the very sensitive BlaR-CTD. This method was used to measure the permeability of beta-lactamase-deficient strains of Enterobacter cloacae and Enterobacter aerogenes to benzylpenicillin, ampicillin, carbenicillin, cefotaxime, aztreonam, and cephacetrile. Except for that of cephacetrile, the permeability coefficients were equal to or below 10(-7) cm/s. For cephacetrile, carbenicillin, and benzylpenicillin, the outer membrane of E. cloacae was 20 to 60 times less permeable than that of Escherichia coli, whereas for cefotaxime, aztreonam, and ampicillin it was, respectively, 400, 1,000, and 700 times less permeable. The permeability coefficient for aztreonam is the lowest ever measured (P = 3.2 X 10(-9) cm/s). Using these values, the MICs for a beta-lactamase-overproducing strain of E. cloacae were successfully predicted, demonstrating the validity of the method.
Disciplines :
Microbiology
Pharmacy, pharmacology & toxicology
Author, co-author :
Lakaye, Bernard ;  Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Biochimie et physiologie humaine et pathologique
Dubus, Alice ;  Université de Liège - ULiège > Département de chimie (sciences) > Département de chimie (sciences)
Joris, Bernard ;  Université de Liège - ULiège > Centre d'ingénierie des protéines
Frère, Jean-Marie ;  Université de Liège - ULiège > Département des sciences de la vie > Département des sciences de la vie
Language :
English
Title :
Method for estimation of low outer membrane permeability to beta-lactam antibiotics
Publication date :
September 2002
Journal title :
Antimicrobial Agents and Chemotherapy
ISSN :
0066-4804
eISSN :
1098-6596
Publisher :
Amer Soc Microbiology, Washington, United States - Washington
Volume :
46
Issue :
9
Pages :
2901-2907
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 17 September 2009

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