scholarly journals Structural determination of a 5-acetamido-3,5,7,9-tetradeoxy-7-(3-hydroxybutyramido)-L-glycero-L-manno-nonulosonic acid-containing homopolysaccharide isolated from Sinorhizobium fredii HH103

1999 ◽  
Vol 342 (3) ◽  
pp. 527-535 ◽  
Author(s):  
Antonio M. GIL-SERRANO ◽  
Miguel A. RODRÍGUEZ-CARVAJAL ◽  
Pilar TEJERO-MATEO ◽  
José L. ESPARTERO ◽  
Manuel MENENDEZ ◽  
...  
Keyword(s):  
Fast Atom Bombardment ◽  
Laser Desorption Ionization ◽  
Glycosidic Linkage ◽  
Methylation Analysis ◽  
Sinorhizobium Fredii ◽  
One Dimensional ◽  
Nonulosonic Acid ◽  
K Antigen ◽  
Fast Atom Bombardment Ms

The structure of a polysaccharide from Sinorhizobium fredii HH103 has been determined. This polysaccharide was isolated by following the protocol for lipopolysaccharide extraction. On the basis of monosaccharide analysis, methylation analysis, fast atom bombardment MS, matrix-assisted laser desorption ionization MS, electron-impact high-resolution MS, one-dimensional 1H-NMR and 13C-NMR and two-dimensional NMR experiments, the structure was shown to consist of a homopolymer of a 3:1 mixture of 5-acetamido-3,5,7,9-tetradeoxy-7-[(R)- and (S)-3-hydroxybutyramido]-L-glycero-L-manno-nonulosonic acid. The sugar residues are attached via a glycosidic linkage to the OH group of the 3-hydroxybutyramido substituent and thus the monomers are linked via both glycosidic and amidic linkages. In contrast with the Sinorhizobium K-antigens previously reported, which are composed of a disaccharide repeating unit, the K-antigen polysacharide of S. fredii HH103 is a homopolysaccharide.

scholarly journals Structural determination of a 5-O-methyl-deaminated neuraminic acid (Kdn)-containing polysaccharide isolated from Sinorhizobium fredii

1998 ◽  
Vol 334 (3) ◽  
pp. 585-594 ◽  
Author(s):  
Antonio M. GIL-SERRANO ◽  
Miguel A. ÍGUEZ-CARVAJAL RODR ◽  
Pilar TEJERO-MATEO ◽  
José L. ESPARTERO ◽  
Jane THOMAS-OATES ◽  
...  
Keyword(s):  
Fast Atom Bombardment ◽  
Wild Type Strain ◽  
Auxotrophic Mutant ◽  
Neuraminic Acid ◽  
Wild Type ◽  
1H And 13C Nmr ◽  
One Dimensional ◽  
In The Wild ◽  
Fast Atom Bombardment Ms

The structure of a polysaccharide from Sinorhizobium frediiSVQ293, a thiamine auxotrophic mutant of S. fredii HH103, has been determined. This polysaccharide was isolated following the protocol for lipopolysaccharide extraction. On the basis of monosaccharide analysis, methylation analysis, fast atom bombardment MS, collision-induced dissociation tandem MS, one-dimensional 1H and 13C NMR and two-dimensional NMR experiments, the structure was shown to consist of the following trisaccharide repeating unit → 2)-α-d-Galp-(1 → 2)-β-d-Ribf-(1 → 9)-α-5-O-Me-Kdnp-(2 →, in which Kdn stands for deaminated neuraminic acid; 25% of the Kdn residues are not methylated. The structure of this polysaccharide is novel and this is the first report of the presence of Kdn in a rhizobial polysaccharide, as well as being the first structure described containing 5-O-Me-Kdn. This Kdn-containing polysaccharide is not present in the wild-type strain HH103, which produces a 3-deoxy-d-manno-2-octulosonic acid (Kdo)-rich polysaccharide. We conclude that it is likely that the appearance of this new Kdn-containing polysaccharide is a consequence of the mutation.

scholarly journals Determination of the structure of a novel acidic oligosaccharide with blood-group activity isolated from bovine submaxillary-gland mucin

1989 ◽  
Vol 260 (2) ◽  
pp. 389-393 ◽  
Author(s):  
S M D'Arcy ◽  
C M Donoghue ◽  
C A M Koeleman ◽  
D H Van den Eijnden ◽  
A V Savage
Keyword(s):  
Blood Group ◽  
Submaxillary Gland ◽  
Two Dimensional ◽  
Methylation Analysis ◽  
One Dimensional ◽  
First Report ◽  
Novel Structure ◽  
Blood Group H

In this study we investigated the structure of an acidic fucose-containing pentasaccharide released from bovine submaxillary-gland mucin by alkaline-borohydride treatment. The structure, determined by a combination of one-dimensional and two-dimensional 1H-n.m.r. spectroscopy at 270 MHz and methylation analysis involving g.l.c.-m.s., was as follows: Fuc alpha(1-2)Gal beta(1-4)GlcNAc beta(1-3)[NeuAc alpha(2-6)]GalNAcol This pentasaccharide is a novel structure and is the first report of a blood-group-H type 2 determinant on a submaxillary-gland mucin.

scholarly journals Structural determination of a 5-acetamido-3,5,7,9-tetradeoxy-7-(3-hydroxybutyramido)-L-glycero-L-manno-nonulosonic acid-containing homopolysaccharide isolated from Sinorhizobium fredii HH103

1999 ◽  
Vol 342 (3) ◽  
pp. 527 ◽  
Author(s):  
Antonio M. GIL-SERRANO ◽  
Miguel A. RODRÍGUEZ-CARVAJAL ◽  
Pilar TEJERO-MATEO ◽  
José L. ESPARTERO ◽  
Manuel MENENDEZ ◽  
...  
Keyword(s):  
Structural Determination ◽  
Sinorhizobium Fredii ◽  
Nonulosonic Acid

scholarly journals Determination of the chemical structure of the capsular polysaccharide of strain B33, a fast-growing soya bean-nodulating bacterium isolated from an arid region of China

2001 ◽  
Vol 357 (2) ◽  
pp. 505-511 ◽  
Author(s):  
Miguel A. RODRÍGUEZ-CARVAJAL ◽  
Pilar TEJERO-MATEO ◽  
José L. ESPARTERO ◽  
José. E. RUIZ-SAINZ ◽  
Ana M. BUENDÍA-CLAVERÍA ◽  
...  
Keyword(s):  
Capsular Polysaccharide ◽  
Soya Bean ◽  
Structural Motif ◽  
Nitrogen Fixing ◽  
1H And 13C Nmr ◽  
One Dimensional ◽  
Fast Growing ◽  
Surface Polysaccharide ◽  
K Antigen

We have determined the structure of a polysaccharide from strain B33, a fast-growing bacterium that forms nitrogen-fixing nodules with Asiatic and American soya bean cultivars. On the basis of monosaccharide analysis, methylation analysis, one-dimensional 1H- and 13C-NMR and two-dimensional NMR experiments, the structure was shown to consist of a polymer having the repeating unit → 6)-4-O-methyl-α-d-Glcp-(1 → 4)-3-O-methyl-β-d-GlcpA-(1 → (where GlcpA is glucopyranuronic acid and Glcp is glucopyranose). Strain B33 produces a K-antigen polysaccharide repeating unit that does not have the structural motif sugar-Kdx [where Kdx is 3-deoxy-d-manno-2-octulosonic acid (Kdo) or a Kdo-related acid] proposed for different Sinorhizobium fredii strains, all of them being effective with Asiatic soya bean cultivars but unable to form nitrogen-fixing nodules with American soya bean cultivars. Instead, it resembles the K-antigen of S. fredii strain HH303 (rhamnose, galacturonic acid)n, which is also effective with both groups of soya bean cultivars. Only the capsular polysaccharide from strains B33 and HH303have monosaccharide components that are also present in the surface polysaccharide of Bradyrhizobium elkanii strains, which consists of a 4-O-methyl-d-glucurono-l-rhamnan.

scholarly journals Structure of the unusual Sinorhizobium fredii HH103 lipopolysaccharide and its role in symbiosis

2020 ◽  
Vol 295 (32) ◽  
pp. 10969-10987 ◽  
Author(s):  
Flaviana Di Lorenzo ◽  
Immacolata Speciale ◽  
Alba Silipo ◽  
Cynthia Alías-Villegas ◽  
Sebastián Acosta-Jurado ◽  
...  
Keyword(s):  
Acid Derivative ◽  
Lipid A ◽  
Core Oligosaccharide ◽  
Sinorhizobium Fredii ◽  
Symbiotic Associations ◽  
Nonulosonic Acid ◽  
Surface Polysaccharides ◽  
Pseudaminic Acid ◽  
Macroptilium Atropurpureum ◽  
K Antigen

Rhizobia are soil bacteria that form important symbiotic associations with legumes, and rhizobial surface polysaccharides, such as K-antigen polysaccharide (KPS) and lipopolysaccharide (LPS), might be important for symbiosis. Previously, we obtained a mutant of Sinorhizobium fredii HH103, rkpA, that does not produce KPS, a homopolysaccharide of a pseudaminic acid derivative, but whose LPS electrophoretic profile was indistinguishable from that of the WT strain. We also previously demonstrated that the HH103 rkpLMNOPQ operon is responsible for 5-acetamido-3,5,7,9-tetradeoxy-7-(3-hydroxybutyramido)-l-glycero-l-manno-nonulosonic acid [Pse5NAc7(3OHBu)] production and is involved in HH103 KPS and LPS biosynthesis and that an HH103 rkpM mutant cannot produce KPS and displays an altered LPS structure. Here, we analyzed the LPS structure of HH103 rkpA, focusing on the carbohydrate portion, and found that it contains a highly heterogeneous lipid A and a peculiar core oligosaccharide composed of an unusually high number of hexuronic acids containing β-configured Pse5NAc7(3OHBu). This pseudaminic acid derivative, in its α-configuration, was the only structural component of the S. fredii HH103 KPS and, to the best of our knowledge, has never been reported from any other rhizobial LPS. We also show that Pse5NAc7(3OHBu) is the complete or partial epitope for a mAb, NB6-228.22, that can recognize the HH103 LPS, but not those of most of the S. fredii strains tested here. We also show that the LPS from HH103 rkpM is identical to that of HH103 rkpA but devoid of any Pse5NAc7(3OHBu) residues. Notably, this rkpM mutant was severely impaired in symbiosis with its host, Macroptilium atropurpureum.

Improved thin-layer chromatographic determination of phospholipids in gastric aspirate from newborns, for assessment of lung maturity.

1988 ◽  
Vol 34 (4) ◽  
pp. 736-738 ◽  
Author(s):  
D Serrano de la Cruz ◽  
E Santillana ◽  
A Mingo ◽  
G Fuenmayor ◽  
A Pantoja ◽  
...  
Keyword(s):  
Thin Layer ◽  
Fetal Lung ◽  
Chromatographic Determination ◽  
Solvent Mixture ◽  
Gastric Aspirate ◽  
One Dimensional ◽  
Acetic Acid Water ◽  
Chromatographic Resolution ◽  
Lung Maturity

Abstract This one-dimensional thin-layer chromatographic method is used for assay of phospholipids in the gastric aspirate of newborns. The solvent mixture (chloroform/hexane/methanol/glacial acetic acid/water, 12/7/4/3/0.3 by vol) completely resolves lecithin, sphingomyelin, phosphatidylinositol, phosphatidylserine, phosphatidylethanolamine, and phosphatidylglycerol. The method is simple, precise, inexpensive, and rapid (chromatographic development takes less than 25 min) and gives high chromatographic resolution. We used this method to determine the lecithin/sphingomyelin densitometric ratio (L/S ratio) and the phosphatidylglycerol percentage in 200 samples of gastric aspirate and found an L/S ratio of 2.5 to be a satisfactory cutoff value for distinguishing fetal lung maturity and immaturity. We confirmed that the presence of phosphatidylglycerol excluded the possibility of respiratory distress.

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