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Journal of Bacteriology, October 1999, p. 5909-5914, Vol. 181, No. 19
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Expression of the Staphylococcus aureus UDP-N-Acetylmuramoyl- L-Alanyl-D-Glutamate:L-Lysine Ligase in Escherichia coli and Effects on Peptidoglycan Biosynthesis and Cell Growth

Dominique Mengin-Lecreulx,^1,* Tim Falla,^2, Didier Blanot,¹ Jean van Heijenoort,¹ David J. Adams,² and Ian Chopra²

Laboratoire des Enveloppes Bactériennes, Centre National de la Recherche Scientifique, Université Paris-Sud, Orsay, France,¹ and Division of Microbiology and Antimicrobial Research Centre, University of Leeds, Leeds LS2 9JT, United Kingdom²

Received 7 May 1999/Accepted 19 July 1999

The monomer units in the Escherichia coli and Staphylococcus aureus cell wall peptidoglycans differ in the nature of the third amino acid in the L-alanyl--D-glutamyl-X-D-alanyl-D-alanine side chain, where X is meso-diaminopimelic acid or L-lysine, respectively. The murE gene from S. aureus encoding the UDP-N-acetylmuramoyl-L-alanyl-D-glutamate: L-lysine ligase was identified and cloned into plasmid vectors. Induction of its overexpression in E. coli rapidly results in abnormal morphological changes and subsequent cell lysis. A reduction of 28% in the peptidoglycan content was observed in induced cells, and analysis of the peptidoglycan composition and structure showed that ca. 50% of the meso-diaminopimelic acid residues were replaced by L-lysine. Lysine was detected in both monomer and dimer fragments, but the acceptor units from the latter contained exclusively meso-diaminopimelic acid, suggesting that no transpeptidation could occur between the -amino group of L-lysine and the -carboxyl group of D-alanine. The overall cross-linking of the macromolecule was only slightly decreased. Detection and analysis of meso-diaminopimelic acid- and L-lysine-containing peptidoglycan precursors confirmed the presence of L-lysine in precursors containing amino acids added after the reaction catalyzed by the MurE ligase and provided additional information about the specificity of the enzymes involved in these latter processes.

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^* Corresponding author. Mailing address: Biochimie Structurale et Cellulaire, EP1088 CNRS, Université Paris-Sud, Bâtiment 430, 91405 Orsay Cedex, France. Phone: 33-1-69156134. Fax: 33-1-69-85-37-15. E-mail: dominique.mengin-lecreulx{at}ebp.u-psud.fr.

Present address: Intrabiotics Pharmaceuticals, Inc., Mountain View, CA 94043.

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Journal of Bacteriology, October 1999, p. 5909-5914, Vol. 181, No. 19
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

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