The wavB gene of Vibrio cholerae and the waaE of Klebsiella pneumoniae codify for a β-1,4-glucosyltransferase involved in the transfer of a glucose residue to the ?-glycero-?-manno-heptose I in the lipopolysaccharide inner core

Abstract

Vibrio cholerae WavB protein showed some similarity to WaaE of Klebsiella pneumoniae and Serratia marcescens. From previous data obtained by us and by chemical analyses of a K. pneumoniae non-polar waaE mutant from strain 889 (08:K69), its lipopolysaccharide (LPS) core structure has recently been elucidated. We demonstrated that WaaE is a beta-1,4-glucosyltransferase involved in the transfer of a glucose residue to the L-glycero-D-manno-heptose I in the LPS inner core. Complementation of this K. pneumoniae non-polar waaE mutant with gene wavB obtained, either from V. cholerae or V. mimicus, showed a full complementation either by chemical studies or by a biological test (susceptibility to non-immune serum). The V. cholerae wavB gene is located in a putative core oligosaccharide (OS) gene cluster and the V. cholerae OS core structure showed the same beta-1,4-glucose residue attached to Hep I as is observed for the K. pneumoniae 889 OS core structure. No other glucose residue is found in the ligosaccharide core structure of K. pneumoniae 889. We concluded that WavB protein is able to perform the same function as WaaE.

A Second Outer-Core Region in Klebsiella pneumoniae Lipopolysaccharide

“…Depending on the K. pneumoniae strain, residues J and K could be H or GalA, and residue P could be H or Hep. Broken lines denote the truncation level for the different core biosynthetic gene mutations (12,(18)(19)(20)32). (B) Type 2 core found in K. pneumoniae 52145 (O1:K2).…”

“…Second, sequence-derived primers were used for further PCR amplification and sequence determination to close the sequenced gaps. Analysis of the data obtained revealed homologues of the reported strain C3 genes encoding L,D-heptose epimerase (hldD), heptosyltransferases I, II, and III (waaC, waaF, and waaQ), 3-deoxy-D-manno-octϩulosonic acid (Kdo) transferase (waaA) (32), inner-core glucosyltransferase (waaE) (18,20), galacturonyltransferase (wabG) (19), N-acetyl-glucosaminyltransferase (wabH) (12), and a gene of unknown function (orf10) similar to E. coli yibD (32) (Fig. 2).…”

The Incorporation of Glucosamine into Enterobacterial Core Lipopolysaccharide

“…1). Furthermore the functions in both inner and outer core biosynthesis of most of the transferases encoded by the wa gene products have been elucidated (7)(8)(9)(10)(11)(12) (Fig. 1).…”

A Second Galacturonic Acid Transferase Is Required for Core Lipopolysaccharide Biosynthesis and Complete Capsule Association with the Cell Surface in Klebsiella pneumoniae

“…(B) The structure of K. pneumoniae type 2 core OS (22). Broken lines denote the truncation level for the different core biosynthetic gene mutations (6, 11-13, 21, 22 1 H nuclear magnetic resonance ( 1 H-NMR) spectra were recorded in D 2 O at 400 MHz with a Bruker DRX 400 Avance spectrometer equipped with a reverse probe in the Fourier Tranform mode at 303 K. 13 C and 1 H chemical shifts were measured in D 2 O using acetone at ␦ values of 2.222 and 31.45 for proton and carbon, respectively, as internal standards. Two-dimensional homo-and heteronuclear experiments (correlated spectroscopy, total correlation spectroscopy, nuclear Overhauser effect spectroscopy, and heteronuclear single-quantum correlation [HSQC]) were performed using standard pulse sequences available in the Bruker software.…”