title: 3-Deoxy-D-manno-octulosonic acid (Kdo) hydrolase identified in Francisella tularensis, Helicobacter pylori, and Legionella pneumophila.
abstract: 3-Deoxy-D-manno-octulosonic acid (Kdo) is an eight-carbon sugar ubiquitous in Gram-negative bacterial lipopolysaccharides (LPS). Although its biosynthesis is well described, no protein has yet been identified as a Kdo hydrolase. However, Kdo hydrolase enzymatic activity has been detected in membranes of Helicobacter pylori and Francisella tularensis and may be responsible for the removal of side-chain Kdo from the LPS core saccharides. We now report the identification of genes encoding a Kdo hydrolase in F. tularensis Schu S4 and live vaccine strain strains, in H. pylori 26695 strain and in Legionella pneumophila Philadelphia 1 strain. We have renamed the genes kdhA for keto-deoxyoctulosonate hydrolase A. Deletion of kdhA abolished Kdo hydrolase activity in membranes of F. tularensis live vaccine strain. The F. tularensis kdhA mutant synthesized a core oligosaccharide containing a Kdo disaccharide with one of the Kdo residues being a terminal side chain. This side-chain Kdo monosaccharide was absent in the wild-type core oligosaccharide. Expression in Escherichia coli of recombinant KdhA from F. tularensis, H. pylori, and L. pneumophila resulted in a reduction of membrane-associated side-chain Kdo. The identification of this previously faceless enzyme will accelerate study of the biosynthetic basis and biologic impact for postbiosynthetic LPS structural modification.
authors.affiliation: Department of Microbiology and Molecular Genetics, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Sabina
authors.initials: S
authors.lastname: Chalabaev
authors.forename: Tae-Hyun
authors.initials: TH
authors.lastname: Kim
authors.forename: Robin
authors.initials: R
authors.lastname: Ross
authors.forename: Alec
authors.initials: A
authors.lastname: Derian
authors.forename: Dennis L
authors.initials: DL
authors.lastname: Kasper
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001428:Bacterial Vaccines / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology / Q000235:genetics; D005604:Francisella tularensis / Q000201:enzymology* / Q000235:genetics; D017353:Gene Deletion; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism*; D016480:Helicobacter pylori / Q000201:enzymology* / Q000235:genetics; D016952:Legionella pneumophila / Q000201:enzymology* / Q000235:genetics; D008070:Lipopolysaccharides / Q000235:genetics / Q000378:metabolism*; D009844:Oligosaccharides / Q000235:genetics / Q000378:metabolism; D013400:Sugar Acids / Q000378:metabolism*; D014613:Vaccines, Attenuated / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D008070:Lipopolysaccharides; D009844:Oligosaccharides; D013400:Sugar Acids; D014613:Vaccines, Attenuated; C002532:2-keto-3-deoxyoctonate; D006026:Glycoside Hydrolases; C053605:KDO alpha-octulopyranoside hydrolase
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