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Journal of Bacteriology, September 2001, p. 5122-5127, Vol. 183, No. 17
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.17.5122-5127.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Identification of the UDP-MurNAc-Pentapeptide:L-Alanine Ligase for Synthesis of Branched Peptidoglycan Precursors in Enterococcus faecalis

Ahmed Bouhss,1,2 Nathalie Josseaume,1 David Allanic,1 Muriel Crouvoisier,2 Laurent Gutmann,1 Jean-Luc Mainardi,1 Dominique Mengin-Lecreulx,2 Jean van Heijenoort,2 and Michel Arthur1,2,*

INSERM E0004-LRMA, UFR Broussais-Hôtel Dieu, Université Paris VI, 75270 Paris,1 and Biochimie Moléculaire et Cellulaire, UMR 8619 CNRS, Université Paris-Sud, 91405 Orsay Cedex,2 France

Received 20 February 2001/Accepted 13 June 2001

Many species of gram-positive bacteria produce branched peptidoglycan precursors resulting from the transfer of various L-amino acids or glycine from amino acyl-tRNA to the varepsilon -amino group of L-lysine. The UDP-MurNAc-pentapeptide:L-alanine ligase and alanyl-tRNA synthetase genes from Enterococcus faecalis were identified, cloned, and overexpressed in Escherichia coli. The purified enzymes were necessary and sufficient for tRNA-dependent addition of L-alanine to UDP-MurNAc-pentapeptide in vitro. The ligase belonged to the Fem family of proteins, which were initially identified genetically as factors essential for methicillin resistance in Staphylococcus aureus.

* Corresponding author. Mailing address: LRMA, Université Paris VI, 15 rue de l'Ecole de Médecine, 75270 Paris Cedex 06, France. Phone: 33 (0)1 43 25 00 33. Fax: 33 (0)1 43 25 68 12. E-mail: michel.arthur{at}bhdc.jussieu.fr.

Journal of Bacteriology, September 2001, p. 5122-5127, Vol. 183, No. 17
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.17.5122-5127.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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