Exopolysaccharide biosynthesis in Lactococcus lactis NIZO B40: functional analysis of the glycosyltransferase genes involved in synthesis of the polysaccharide backbone.
We used homologous and heterologous expression of the glycosyltransferase genes of the Lactococcus lactis NIZO B40 eps gene cluster to determine the activity and substrate specificities of the encoded enzymes and established the order of assembly of the trisaccharide backbone of the exopolysaccharide repeating unit. EpsD links glucose-1-phosphate from UDP-glucose to a lipid carrier, EpsE and EpsF link glucose from UDP-glucose to lipid-linked glucose, and EpsG links galactose from UDP-galactose to lipid-linked cellobiose. Furthermore, EpsJ appeared to be involved in EPS biosynthesis as a galactosyl phosphotransferase or an enzyme which releases the backbone oligosaccharide from the lipid carrier.
van Kranenburg
R
R
Microbial Ingredients Section, NIZO Food Research, 6710 BA Ede, The Netherlands.kranenbu@nizo.nl
van Swam
I I
II
Marugg
J D
JD
Kleerebezem
M
M
de Vos
W M
WM
D001483:Base Sequence; D002240:Carbohydrate Sequence; D002874:Chromosome Mapping; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D016695:Glycosyltransferases / Q000235:genetics* / Q000378:metabolism*; D013294:Lactococcus lactis / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D005810:Multigene Family; D016133:Polymerase Chain Reaction; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis* / Q000737:chemistry; D013379:Substrate Specificity
D017931:DNA Primers; D011135:Polysaccharides, Bacterial; D016695:Glycosyltransferases