TY - JOUR
T1 - The in vitro biosynthesis of the exopolysaccharide produced by Rhizobium leguminosarum bv. trifolii, strain NA 30.
AU - Bossio, J. C.
AU - Semino, C. E.
AU - Iñón de Iannino, N.
AU - Dankert, M. A.
PY - 1996/7
Y1 - 1996/7
N2 - Rhizobium leguminosarum bv. trifolii, strain NA 30 nodulates both red (Trifolium pratense) and white (T. repens) clover and produces an acidic exopolysaccharide (EPS) containing glucose, galactose, glucuronic acid, acetate and ketalpyruvate residues in a 5:1:2:1:2 molar ratio. The in vitro synthesis of this EPS as well as the characterization of five structurally related lipid linked oligosaccharides is described employing EDTA treated cells as enzyme preparation and 14C-labelled UDP-Glc, UDP-GlcA, Acetyl CoA and phosphoenol pyruvate (PEP) and doubly labelled 32P UDP-14C-Glc as precursors. The lipidic derivatives, extracted with chloroform, methanol, water (1:2:0,3) had the properties expected for prenyl-diphospho-sugars, as judged by the pattern of labelling, DEAE cellulose column chromatography, catalytic reduction and acid lability, etc. The sugar moieties of these phosphoprenyl derivatives were identified as the acetylated octasaccharide repeating unit, its mono- and di-ketalpyruvate derivatives and two trisaccharides, one of them acetylated, on the basis of specific labelling, gel filtration, paper electrophoresis and chromatography, TLC, permethylation, etc. In vitro polymer synthesis was greatly increased when electroporated cells were substituted for EDTA treated cells as enzyme system.
AB - Rhizobium leguminosarum bv. trifolii, strain NA 30 nodulates both red (Trifolium pratense) and white (T. repens) clover and produces an acidic exopolysaccharide (EPS) containing glucose, galactose, glucuronic acid, acetate and ketalpyruvate residues in a 5:1:2:1:2 molar ratio. The in vitro synthesis of this EPS as well as the characterization of five structurally related lipid linked oligosaccharides is described employing EDTA treated cells as enzyme preparation and 14C-labelled UDP-Glc, UDP-GlcA, Acetyl CoA and phosphoenol pyruvate (PEP) and doubly labelled 32P UDP-14C-Glc as precursors. The lipidic derivatives, extracted with chloroform, methanol, water (1:2:0,3) had the properties expected for prenyl-diphospho-sugars, as judged by the pattern of labelling, DEAE cellulose column chromatography, catalytic reduction and acid lability, etc. The sugar moieties of these phosphoprenyl derivatives were identified as the acetylated octasaccharide repeating unit, its mono- and di-ketalpyruvate derivatives and two trisaccharides, one of them acetylated, on the basis of specific labelling, gel filtration, paper electrophoresis and chromatography, TLC, permethylation, etc. In vitro polymer synthesis was greatly increased when electroporated cells were substituted for EDTA treated cells as enzyme system.
KW - EDTA treated cells
KW - Rhizobium leguminosarum bv trifolii
KW - Biosynthesis
KW - Electroporated cells
KW - Exopolysaccharide
KW - Prenyl-phospho-sugars
UR - http://www.scopus.com/inward/record.url?scp=0030178604&partnerID=8YFLogxK
UR - https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=pure_univeritat_ramon_llull&SrcAuth=WosAPI&KeyUT=WOS:A1996UY06300023&DestLinkType=FullRecord&DestApp=WOS_CPL
M3 - Article
C2 - 8832106
AN - SCOPUS:0030178604
SN - 0145-5680
VL - 42
SP - 737
EP - 758
JO - Cellular and molecular biology (Noisy-le-Grand, France)
JF - Cellular and molecular biology (Noisy-le-Grand, France)
IS - 5
ER -