Open Access Research article

Deletion of the glycosyltransferase bgsB of Enterococcus faecalis leads to a complete loss of glycolipids from the cell membrane and to impaired biofilm formation

Christian Theilacker1*, Irina Sava1, Patricia Sanchez-Carballo2, Yinyin Bao1, Andrea Kropec1, Elisabeth Grohmann1, Otto Holst2 and Johannes Huebner1

Author Affiliations

1 Center for Infectious Diseases and Travel Medicine, University Medical Center Freiburg, Germany

2 Division of Structural Biochemistry, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Germany

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BMC Microbiology 2011, 11:67  doi:10.1186/1471-2180-11-67

Published: 6 April 2011

Additional files

Additional file 1:

Transmission electron microscopy of E. faecalis strains. E. faecalis 12030 wild type (A) and 12030ΔbgsB (B). Bar represents 500 nm.

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Additional file 2:

Autolysis and opsonization of E. faecalis 12030ΔbgsB. A Spontaneous bacterial autolysis. Cells were grown to mid-log phase, resuspended in 10 mM sodium phosphate buffer containing 5% Triton X-100 and the decrease of the OD 600 at 30°C was recorded over time. B Bacterial killing in vitro after 90 min in the presence of 6.5% rabbit complement (white bar), 2 × 107 human PMN plus complement (gray bar) and rabbit antiserum raised against whole bacterial cells (serum dilution 1:2500) plus PMN and complement (black bar). Bars represent means ± SEM.

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Additional file 3:

Characterization of E. faecalis ΔbgsB cell walls. A Thin-layer chromatography of cell membrane total lipid extracts of E. faecalis 12030 wild type (lane 1 and 4), 12030ΔbgsB (lane 2 and 5), 12030ΔbgsA (lane 3 and 6). TLC plates were developed using a solvent system of CHCl3/MeOH/H20 (65:25:4, v/v/v). Staining lane 1 - 3 molybdenum blue, lane 4 - 6 ninhydrin. B SDS PAGE of bacterial whole protein extracts. The material was extracted by disrupting the cells with glass-beads, boiling in Laemmli buffer, separated by 4-12% Bis-Tris gels and stained with Coomassie blue.

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Additional file 4:

Minimal bactericial concentration of E. faecalis strains against antimicrobial peptides. Concentrations are expressed as μg/ml.

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