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Journal of Bacteriology, January 2002, p. 363-369, Vol. 184, No. 2
0021-9193/01/$04.00+0     DOI: 10.1128/JB.184.2.363-369.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Biosynthesis Pathway of ADP-L-glycero-ß-D-manno-Heptose in Escherichia coli

Bernd Kneidinger,1 Cristina Marolda,2 Michael Graninger,1 Alla Zamyatina,3 Fiona McArthur,2 Paul Kosma,3 Miguel A. Valvano,2 and Paul Messner1*

Zentrum für Ultrastrukturforschung und Ludwig Boltzmann-Institut für Molekulare Nanotechnologie, A-1180 Vienna,1 Institut für Chemie, Universität für Bodenkultur Wien, A-1190 Vienna, Austria,3 Department of Microbiology and Immunology, University of Western Ontario, London, Ontario N6A 5C1, Canada2

Received 21 August 2001/ Accepted 17 October 2001

The steps involved in the biosynthesis of the ADP-L-glycero-ß-D-manno-heptose (ADP-L-ß-D-heptose) precursor of the inner core lipopolysaccharide (LPS) have not been completely elucidated. In this work, we have purified the enzymes involved in catalyzing the intermediate steps leading to the synthesis of ADP-D-ß-D-heptose and have biochemically characterized the reaction products by high-performance anion-exchange chromatography. We have also constructed a deletion in a novel gene, gmhB (formerly yaeD), which results in the formation of an altered LPS core. This mutation confirms that the GmhB protein is required for the formation of ADP-D-ß-D-heptose. Our results demonstrate that the synthesis of ADP-D-ß-D-heptose in Escherichia coli requires three proteins, GmhA (sedoheptulose 7-phosphate isomerase), HldE (bifunctional D-ß-D-heptose 7-phosphate kinase/D-ß-D-heptose 1-phosphate adenylyltransferase), and GmhB (D,D-heptose 1,7-bisphosphate phosphatase), as well as ATP and the ketose phosphate precursor sedoheptulose 7-phosphate. A previously characterized epimerase, formerly named WaaD (RfaD) and now renamed HldD, completes the pathway to form the ADP-L-ß-D-heptose precursor utilized in the assembly of inner core LPS.

* Corresponding author. Mailing address for Paul Messner: Zentrum für Ultrastrukturforschung, Universität für Bodenkultur Wien, Gregor-Mendel-Str. 33, A-1180 Vienna, Austria. Phone: 43-1-47654-2202. Fax: 43-1-4789112. E-mail: pmessner{at}edv1.boku.ac.at. Mailing address for Miguel A. Valvano: Department of Microbiology and Immunology, University of Western Ontario, London, Ontario N6A 5C1, Canada. Phone: (519) 661-3996. Fax: (519) 661-3499. E-mail: mvalvano{at}uwo.ca.

Journal of Bacteriology, January 2002, p. 363-369, Vol. 184, No. 2
0021-9193/01/$04.00+0     DOI: 10.1128/JB.184.2.363-369.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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