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Infection and Immunity, July 2006, p. 4164-4171, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00111-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Alanine Esters of Enterococcal Lipoteichoic Acid Play a Role in Biofilm Formation and Resistance to Antimicrobial Peptides

Francesca Fabretti,1,2 Christian Theilacker,1 Lucilla Baldassarri,2 Zbigniew Kaczynski,3,4 Andrea Kropec,1 Otto Holst,3 and Johannes Huebner1*

Division of Infectious Diseases, Department of Medicine, Freiburg University Hospital, Freiburg, Germany,1 Dipartimento di Malattie Infettive, Parassitarie ed Immunomediate-Istituto Superiore di Sanitá, Rome, Italy,2 Division of Structural Biochemistry, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany,3 Department of Chemistry, University of Gdansk, Gdansk, Poland4

Received 22 January 2006/ Returned for modification 2 March 2006/ Accepted 20 April 2006

Enterococcus faecalis is among the predominant causes of nosocomial infections. Surface molecules like D-alanine lipoteichoic acid (LTA) perform several functions in gram-positive bacteria, such as maintenance of cationic homeostasis and modulation of autolytic activities. The aim of the present study was to evaluate the effect of D-alanine esters of teichoic acids on biofilm production and adhesion, autolysis, antimicrobial peptide sensitivity, and opsonic killing. A deletion mutant of the dltA gene was created in a clinical E. faecalis isolate. The absence of D-alanine in the LTA of the dltA deletion mutant was confirmed by nuclear magnetic resonance spectroscopy. The wild-type strain and the deletion mutant did not show any significant differences in growth curve, morphology, or autolysis. However, the mutant produced significantly less biofilm when grown in the presence of 1% glucose (51.1% compared to that of the wild type); adhesion to eukaryotic cells was diminished. The mutant absorbed 71.1% of the opsonic antibodies, while absorption with the wild type resulted in a 93.2% reduction in killing. Sensitivity to several cationic antimicrobial peptides (polymyxin B, colistin, and nisin) was considerably increased in the mutant strain, confirming similar results from other studies of gram-positive bacteria. Our data suggest that the absence of D-alanine in LTA plays a role in environmental interactions, probably by modulating the net negative charge of the bacterial cell surface, and therefore it may be involved in the pathogenesis of this organism.

* Corresponding author. Mailing address: Division of Infectious Diseases, Hugstetter Str. 55, 79106 Freiburg, Germany. Phone: 49-761-270-1828. Fax: 49-761-270-1820. E-mail: johannes.huebner{at}uniklinik-freiburg.de.

Editor: J. N. Weiser

Infection and Immunity, July 2006, p. 4164-4171, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00111-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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