Journal of Bacteriology, October 1999, p. 6354-6360, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Nestlé Research Center, Nestec Ltd., Vers-chez-les-Blanc, 1000 Lausanne 26, Switzerland
Received 3 May 1999/Accepted 26 July 1999
Streptococcus thermophilus Sfi6 produces a texturizing
exopolysaccharide (EPS) consisting of a
3)[
-D-Galp-(1
6)]-
-D-Glcp-(1
3)-
-D-GalpNAc-(1
3)-
-D-Galp-(1
repeating
unit. We previously identified and analyzed a 14.5-kb gene cluster from
S. thermophilus Sfi6 consisting of 13 genes responsible for
its EPS production. Within this gene cluster, we found a central region
of genes (epsE, epsF, epsG, and
epsI) that showed similarity to glycosyltransferases. In
this study, we investigated the sugar specificity of these enzymes.
EpsE catalyzes the first step in the biosynthesis of the EPS repeating
unit. It exhibits phosphogalactosyltransferase activity and transfers galactose onto the lipophilic carrier. The second step is fulfilled by
EpsG, which transfers an
-N-acetylgalactosamine onto the
first
-galactoside. The activity of EpsF was determined by
characterizing the EPS produced by an S. thermophilus epsF
deletion mutant. This EPS consisted of the monosaccharides Gal, Glc,
and GalNAc in an approximately equimolar ratio, thus suggesting that
epsF codes for the branching galactosyltransferase.
epsI probably codes for the
-1,3-glucosyltransferase,
since it is the only glycosyltransferase to which no gene has been
assigned and it exhibits similarity to other
-glycosyltransferases.
EpsE shows the conserved features of phosphoglycosyltransferases,
whereas EpsF and EpsG exhibit the primary structure of
-glycosyltransferases, belonging to glycosyltransferase family 4, whose members are conserved in all major phylogenetic lineages,
including the Archaea and Eukaryota.
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