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Journal of Bacteriology, May 1999, p. 2726-2732, Vol. 181, No. 9
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Identification and Characterization of a New Porin Gene of Klebsiella pneumoniae: Its Role in beta -Lactam Antibiotic Resistance

Antonio Doménech-Sánchez,1 Santiago Hernández-Allés,1 Luis Martínez-Martínez,2 Vicente J. Benedí,1 and Sebastián Albertí1,*

Área de Microbiología, Departamento de Biología, Universidad de las Islas Baleares, and IMEDEA (CSIC-UIB), Palma de Mallorca,1 and Departamento de Microbiología, Universidad de Sevilla, Seville,2 Spain

Received 21 December 1998/Accepted 19 February 1999

Klebsiella pneumoniae porin genes were analyzed to detect mutations accounting for the porin deficiency observed in many beta -lactam-resistant strains. PCR and Southern blot analysis revealed the existence of a third porin gene in addition to the OmpK36 and OmpK35 porin genes previously described. This new porin gene was designated ompK37 and is present in all of the clinical isolates tested. The OmpK37 porin gene was cloned, sequenced, and overexpressed in Escherichia coli. In contrast to that of the major porins, OmpK37 porin expression was only detectable by Western blot analysis in porin-deficient beta -lactam-resistant strains, suggesting strong down regulation under standard laboratory conditions. Functional characterization suggested a narrower pore for the OmpK37 porin than for K. pneumoniae porins OmpK36 and OmpK35. This correlated with the susceptibility to certain beta -lactam antibiotics, since a K. pneumoniae strain expressing porin OmpK37, but not porin OmpK36 or OmpK35, was less susceptible to beta -lactam antibiotics than the same strain expressing either porin OmpK36 or OmpK35.

* Corresponding author. Mailing address: UIB-Microbiología, Carretera de Valldemosa Km. 7.5, 07071-Palma de Mallorca, Spain. Phone: 34-971-173335. Fax: 34-971-173184. E-mail: dbasas3{at}ps.uib.es.

Journal of Bacteriology, May 1999, p. 2726-2732, Vol. 181, No. 9
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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