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PMID: 8626297 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Identification and characterization of the eps (Exopolysaccharide) gene cluster from Streptococcus thermophilus Sfi6.

Journal of bacteriology ·Vol. 178 ·No. 6 ·1996-03-00 ·Pages 1680-90

Stingele F, Neeser JR, Mollet B

Abstract

We report the identification and characterization of the eps gene cluster of Streptococcus thermophilus Sfi6 required for exopolysaccharide (EPS) synthesis. This report is the first genetic work concerning EPS production in a food microorganism. The EPS secreted by this strain consists of the following tetrasaccharide repeating unit:-->3)-beta-D-Galp-(1-->3)-[alpha-D-Galp-(1-->6)]-beta-D- D-Galp-(1-->3)-alpha-D-Galp-D-GalpNAc-(1-->. The genetic locus The genetic locus was identified by Tn916 mutagenesis in combination with a plate assay to identify Eps mutants. Sequence analysis of the gene region, which was obtained from subclones of a genomic library of Sfi6, revealed a 15.25-kb region encoding 15 open reading frames. EPS expression in the non-EPS-producing heterologous host, Lactococcus lactis MG1363, showed that within the 15.25-kb region, a region with a size of 14.52 kb encoding the 13 genes epsA to epsM was capable of directing EPS synthesis and secretion in this host. Homology searches of the predicted proteins in the Swiss-Prot database revealed high homology (40 to 68% identity) for epsA, B, C, D, and E and the genes involved in capsule synthesis in Streptococcus pneumoniae and Streptococcus agalactiae. Moderate to low homology (37 to 18% identity) was detected for epsB, D, F, and H and the genes involved in capsule synthesis in Staphylococcus aureus for epsC, D, and E and the genes involved in exopolysaccharide I (EPSI) synthesis in Rhizobium meliloti for epsC to epsJ and the genes involved in lipopolysaccharide synthesis in members of the Enterobacteriaceae, and finally for eps K and lipB of Neisseria meningitidis. Genes (epsJ, epsL, and epsM) for which the predicted proteins showed little or no homology with proteins in the Swiss-Prot database were shown to be involved in EPS synthesis by single-crossover gene disruption experiments.

MeSH Terms
Bacterial Proteins/genetics Base Composition Base Sequence Carbohydrate Sequence Chromosome Mapping Conserved Sequence DNA Transposable Elements Genes, Bacterial Genomic Library Lactococcus lactis/genetics Molecular Sequence Data Multigene Family Mutagenesis, Insertional Polysaccharides, Bacterial/biosynthesis Recombinant Proteins Sequence Analysis, DNA Sequence Homology, Amino Acid Streptococcus/cytology,genetics
Chemicals
Bacterial Proteins DNA Transposable Elements Polysaccharides, Bacterial Recombinant Proteins exopolysaccharide, Streptococcus
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Stingele F
Nestlé Research Center, Nestec Ltd., Lausanne, Switzerland.
Neeser J R
Mollet B
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.178.6.1680-1690.1996
Published
1996-03-00
Pages
1680-90
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC177854
Subset
IM
Databases
GENBANK
U40830
External Links
PubMed source DOI: 10.1128/jb.178.6.1680-1690.1996 PMC Full Text View journal details
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