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L Kenne

Publications and source records attributed to L Kenne.

At least 55 records · Page 3Linked to original sources

Structural studies of the extracellular polysaccharide from Butyrivibrio fibrisolvens strain X6C61.

The capsular polysaccharide from Butyrivibrio fibrisolvens strain X6C61 has been investigated using NMR spectroscopy, mass spectrometry, methylation analysis, and partial acid hydrolysis as the main methods. The polysaccharide is composed of hexasaccharide repeating units having the following structure. [formula: see text] The polysaccharide also contains O-acetyl groups, of which approximately 70% are substituted to O-3 of the beta-D-Glc pA residue.

Bacterial Capsules↗

Structural studies of the Vibrio mimicus W-26768 O-antigen polysaccharide.

The structure of the Vibrio mimicus W-26768 O-antigen polysaccharide has been investigated by sugar and methylation analyses, Smith degradation, and NMR spectroscopy. It is proposed that it is composed of chains of beta-(1-->4)-linked 3,6-dideoxy-3-[(R)-3-hydroxybutyramido]-D-glucopyranosyl residues (D-Qui(p)3NR), approximately 60% of which are substituted in the 2-position with 2-acetamido-2-deoxy-alpha-D-galactopyranosyl groups. The polysaccharide does not seem to be composed of oligosaccharide repeating units but has a less regular structure, schematically indicated below. [formula: see text]

Carbohydrate Conformation↗

Structural studies of Vibrio fluvialis M-940 O-antigen polysaccharide.

The structure of the Vibrio fluvialis M-940 O-antigen polysaccharide has been investigated by sugar and methylation analyses, specific degradations, NMR spectroscopy, and mass spectrometry. It is proposed that it consists of a heptasaccharide unit having the following structure. alpha-L-Rhap-(1-->2)-alpha-L-Fucp-(1-->2)-alpha-D-Galp-(1--> 2)-alpha-L-Fucp- (1-->3)-beta-D-GlcpA-(1-->4)-alpha-L-Rhap-(1-->3)-beta-D-Glc pNAc-(1--> The heptasaccharide is most probably linked to the 3-position of an alpha-D-galactopyranosyl residue in the core.

Carbohydrate Conformation↗

Alpha-1,4-glucan lyase, a new class of starch/glycogen degrading enzyme. I. Efficient purification and characterization from red seaweeds.

This study presents the first purification and characterization of an alpha-1,4-glucan lyase. The enzyme degraded alpha-1,4-glucan to produce 1,5-anhydrofructose. A simple and efficient purification procedure has been developed and the enzyme has been purified to homogeneity from two red seaweeds Gracilariopsis lemaneiformis and Gracilaria verrucosa. alpha-1,4-Glucan lyase was apparently a single polypeptide as a molecular weight of 111,000 was observed in SDS-gel electrophoresis, and 98,000 by gel filtration chromatography on Sephacryl S-200. Amino acid composition analysis of the enzyme showed high amounts of Asp/Asn, Gly and Glu/Gln. The isoelectric point of the enzyme was 3.9, as revealed by isoelectrofocusing. The concentrations of maltotriose, maltose and amylopectin that yield half of the maximum activity were 798 micrograms ml-1 (1.58 mM), 1,418 micrograms ml-1 (4.14 mM) and 1,600 micrograms ml-1, respectively. alpha-1,4-Glucan lyase exhibited a wide pH optimum range from pH 2.5 to 7.0 for maltose and from pH 3.5 to 7.5 for amylopectin. The optimal temperature for activity of the algal lyase was 50 degrees C when maltose or amylopectin was used as a substrate under the assay conditions. The Arrhenius activation energies were 45.8 and 44.0 kJ mol-1 for maltose and amylopectin as substrate, respectively. Only one form of alpha-1,4-glucan lyase was found in cell-free extracts of the two red seaweeds.

Amino Acids↗

Analysis of 7-substituted sialic acid in some enterobacterial lipopolysaccharides.

Sialic acid-containing lipopolysaccharides (LPS) were isolated from six bacterial strains of the family Enterobacteriaceae. Sialic acid was released from permethylated LPS by methanolysis, and partially O-methylated N-acetyl-N-methyl-neuraminic acid methyl ester methyl glycosides were analyzed by gas-liquid chromatography-electron ionization mass spectrometry. It was proved that all LPS contain N-acetylneuraminic acid (NeuAc). The occurrence of 7-substituted NeuAc in Escherichia coli serotypes O24 and O56 and in Citrobacter freundii O37 LPS was documented. The LPS preparations also contained terminal NeuAc. LPS of E. coli O104 had exclusively 4-substituted sialic acid. The remaining LPS studied, namely, from Salmonella toucra O48 and Hafnia alvei 2, had 4-linked and terminally localized NeuAc residues.

Enterobacteriaceae↗

A 1H and 13C NMR study of oligosaccharides from human milk. Application of the computer program CASPER.

Several oligosaccharides from human milk, containing vicinally branched residues, have been analysed with respect to induced NMR chemical shift changes that originate from the branching. Two types of branching were investigated: (i) linear oligosaccharides with a 2-linked residue, which thus becomes vicinally 1,2-disubstituted, and (ii) oligosaccharides with either 2,3- or 3,4-branching. It could be concluded that, in 13C NMR spectra of the first type, for which only moderately sized induced changes (< 2 ppm) had been observed previously, large (> 5 ppm) changes are also present. For 2,3- and 3,4-branching, changes similar to those observed earlier were found. In 1H NMR spectra, significant induced shifts for signals from anomeric, aglyconic, and H-5 protons were observed. For most trisaccharides, a unique set of values for the chemical shift differences was found, thus making it suitable to use them for characterisation of substitution patterns in the analysis with the computer program CASPER.

Carbohydrate Conformation↗

Synthesis and conformational and NMR studies of alpha-D-mannopyranosyl and alpha-D-mannopyranosyl-(1----2)-alpha-D-mannopyranosyl linked to L-serine and L-threonine.

alpha-D-Mannopyranosyl and alpha-D-mannopyranosyl-(1----2)-alpha-D- mannopyranosyl linked to L-serine and L-threonine have been synthesised as model substances for the linkage region in certain O-linked glycoproteins. Metropolis Monte Carlo simulations were performed with a modified version of the GESA program, to yield theoretical NOEs and interatomic distances as ensemble-average values, and these were compared with results from steady-state NOE experiments. The NOEs were determined as ensemble-average and as global minimum values. NMR chemical shift differences, obtained for signals of the glycopeptides relative to those of the respective monomers, were interpreted in terms of short inter-residue atomic distances as found within the global minima, and on the basis of averaged distances derived from Monte Carlo simulations.

Carbohydrate Conformation↗

Computer-assisted structural analysis of oligosaccharides using CASPER.

The computer program CASPER for structural analysis has been tested on some oligosaccharides. The program is shown to predict the correct structure of five linear or branched tri- to hexasaccharides using information on components and linkage positions and NMR chemical shifts. Theoligosaccharides are either reducting or methyl glycosides.

Carbohydrate Conformation↗

Mapping of the binding specificity for five monoclonal antibodies recognizing 3-deoxy-D-manno-octulosonic acid in bacterial lipopolysaccharides.

Mouse mAb were produced against the deep rough strains Salmonella minnesota R 595, Salmonella typhimurium SL 1102, and Escherichia coli D21f2 and screened by enzyme immunoassay against LPS of several chemotypes. Five antibodies were selected for their ability to bind to chemotype deep rough (Re) LPS which has two 3-deoxy-D-manno-octulosonic acid (Kdo) residues in its nonreducing end. Structurally verified oligosaccharides isolated from rough LPS and synthetic analogues of Kdo were used in an enzyme immunoassay inhibition test to determine the binding epitopes for the antibodies. According to their specificities, the antibodies could be divided into three groups. For two of the groups, the recognized structure was the alpha-Kdo (2----4) Kdo disaccharide and for one group the alpha-Kdo (2----4) alpha-Kdo beta-D-GlcN (1----6) alpha-D-GlcN tetrasaccharide, representing a partial structure of the Re LPS. Inhibition studies with synthetic analogues of Kdo showed that the anomeric configuration and the free carboxyl group of the Kdo residue are important features for antibody binding. Changes in the C-1 to C-6 region of the Kdo molecule do influence the antibody recognition considerably whereas changes in the exocyclic C-7 to C-8 region are of secondary importance. Calculation of the conformation of the inner core region showed that the alpha-Kdo (2----4) alpha-Kdo (2---- disaccharide was free and accessible in chemotype Re LPS, but that linkage of a L-glycero-D-manno-heptose to O-5 of the subterminal Kdo both changes the conformation of the Kdo-disaccharide and covers it thereby making it less accessible.

Animals↗

Structural studies of the extracellular polysaccharide elaborated by Azotobacter vinelandii strain 1484.

The structure of the extracellular polysaccharide from Azotobacter vinelandii strain 1484 has been investigated, specific degradations and n.m.r. spectroscopy being the main methods used. It is concluded that the polysaccharide is composed of tetrasaccharide repeating-units having the following structure, [sequence: see text] in which Sug is 3-deoxy-D-threo-hexulosonic acid. The polysaccharide also contains a non-stoichiometric amount of O-acetyl groups, distributed over at least two positions.

Azotobacter↗

CASPER: a computer program used for structural analysis of carbohydrates.

The computer program CASPER and its algorithms are described. The program is aimed at facilitating the determination of structures of oligosaccharides and regular polysaccharides, requiring as input either the one-dimensional 1H or 13C NMR spectrum or the 2D C,H-correlation NMR spectrum together with information on components and linkages. The databases, the method of simulating spectra, options of the program, and techniques for faster calculations are described as well as an example of a structural determination.

Algorithms↗

Structural studies of the capsular polysaccharide from Aerococcus viridans var. homari.

The capsular polysaccharide from Aerococcus viridans var. homari has been investigated, using n.m.r. spectroscopy, methylation analysis, and specific degradations as the main methods. The polysaccharide is composed of tetrasaccharide repeating-units having the following structure. (Formula; see text) In this structure, D-QuiN stands for 2-amino-2,6-dideoxy-D-glucose (quinovosamine). Two of the three acidic sugars found, namely, L-altruronic acid and 4-O-[(S)-1-carboxyethyl]-D-glucose, have not been found in any other natural source. As evident from the n.m.r. spectra, the L-altruronic acid is not present in the 1C4 conformation, but flips to a conformation close to this on carboxyl reduction.

Carbohydrate Sequence↗

A method for the microanalysis of hexoses in glycoproteins.

A procedure has been developed by which less than micrograms quantities of hexoses in glycoproteins can be determined by g.l.c.-m.s. with selected-ion monitoring of the alditol acetates derived from the sugars released by acid hydrolysis. The effectiveness of the method was demonstrated by determination of the hexose and hexosamine composition of 5-micrograms samples of antithrombin III and von Willebrand factor, respectively.

Antithrombin III↗