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Originally published In Press as doi:10.1074/jbc.M502104200 on May 18, 2005
J. Biol. Chem., Vol. 280, Issue 28, 26573-26585, July 15, 2005
The Crucial Role of Trehalose and Structurally Related Oligosaccharides in the Biosynthesis and Transfer of Mycolic Acids in Corynebacterineae*
Marielle Tropis ,
Xavier Meniche ,
Andreas Wolf¶,
Henrike Gebhardt¶,
Sergey Strelkov¶,
Mohamed Chami||,
Dietmar Schomburg¶,
Reinhard Krämer¶,
Susanne Morbach¶, and
Mamadou Daffé **
From the
Department of Molecular Mechanisms of Mycobacterial Infections, Institut de Pharmacologie et Biologie Structurale (UMR 5089 du CNRS et de l'Université Paul Sabatier) 205, route de Narbonne, 31077 Toulouse cedex 04, France, ¶Institut für Biochemie der Universität zu Köln, Zülpicher Strasse 47, 50674 Köln, Germany, and ||M. E. Müller Institute for Structural Biology, Biozentrum, University of Basel, Klingelbergstrasse 70 CH-4056, Basel, Switzerland
Trehalose ( -D-glucopyranosyl- '-D-glucopyranoside) is essential for the growth of the human pathogen Mycobacterium tuberculosis but not for the viability of the phylogenetically related corynebacteria. To determine the role of trehalose in the physiology of these bacteria, the so-called Corynebacterineae, mutant strains of Corynebacterium glutamicum unable to synthesize trehalose due to the knock-out of the genes of the three pathways of trehalose biosynthesis, were biochemically analyzed. We demonstrated that the synthesis of trehalose under standard conditions is a prerequisite for the production of mycolates, major and structurally important constituents of the cell envelope of Corynebacterineae. Consistently, the trehalose-less cells also lack the cell wall fracture plane that typifies mycolate-containing bacteria. Importantly, however, the mutants were able to synthesize mycolates when grown on glucose, maltose, and maltotriose but not on other carbon sources known to be used for the production of internal glucose phosphate such as fructose, acetate, and pyruvate. The mycoloyl residues synthesized by the mutants grown on -D-glucopyranosyl-containing oligosaccharides were transferred both onto the cell wall and free sugar acceptors. A combination of chemical analytical approaches showed that the newly synthesized glycolipids consisted of 1 mol of mycolate located on carbon 6 of the non reducing glucopyranosyl unit. Additionally, experiments with radioactively labeled trehalose showed that the transfer of mycoloyl residues onto sugars occurs outside the plasma membrane. Finally, and in contradiction to published data, we demonstrated that trehalose 6-phosphate has no impact on mycolate synthesis in vivo.
Received for publication, February 24, 2005
, and in revised form, May 18, 2005.
* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Both authors contributed equally to the work.
** To whom correspondence should be addressed. Tel.: 33-561-175-569; Fax: 33-561-175-580; E-mail: mamadou.daffe{at}ipbs.fr.

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Copyright © 2005 by the American Society for Biochemistry and Molecular Biology.
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