Structure and biodegradation of the polysaccharide components of egg masses isolated from snails of an Ampullarius species

A polysaccharide preparation, isolated from egg masses deposited by snails of an Ampullarius species, was purified via precipitation with Cetavlon in the presence of sodium borate, and found to contain d-galactose and a smaller proportion of d-glucose, and to have two components with sedimentation coefficients of 10S and 40S. A polysaccharide, isolated from freshly laid egg masses, was highly branched and was shown to contain nonreducing α-d-glucopyranosyl and β-d-galactopyranosyl end-groups, and 3,6-di-O-substituted β-d-galactopyranosyl residues. One or more of the polysaccharide components was a d-glucopyrano-d-galactopyranan with non-reducing α-d-glucopyranosyl end-groups (1→4)-linked to β-d-galactopyranosyl residues. The polysaccharide preparations, obtained from freshly laid egg masses and from those that were left for 10 and 15 days after being laid, were structurally different from each other. With the passage of time, progressive diminution of the 10S component and the proportion of d-glucose in the polysaccharide took place, suggesting that each constituent was consumed preferentially by the snail embryos as an energy source.     

The structure of a dextran produced by Leuconostoc mesenteroides NRRL B-1397: the linkages and length of the branches

The structure of a dextran produced by Leuconostoc mesenteroides NRRL B-1397 has been investigated in relation to its immunological properties. The methylated dextran yielded on acid hydrolysis 2,3,4,6-tetra-, 2,3,4-tri-, 3,4,-di-, and 2,4-di-O-methyl-d-glucose, in the molar ratio of 1.0:3.1:0.7:0.2, together with a trace of 2,4,6-tri-O-methyl-dglucose, indicating that the branches occur mainly at O-2 and the remainder at O-3. A carboxyl-dextran, obtained by catalytic oxidation of the dextran to convert the terminal, non-reducing d-glucose residues d-glucuronic acid residues, was partially hydrolyzed with acid. Fractionation gave 2-O-(α-d-glucopyranosyluronic acid) d-glucose (major), 6-O-(α-d-glucopyranosyluronic acid)-d-glucose, and mixtures of aldotri-, aldotetra-, and aldopentaouronic acid that contain both (1 → 6)- and (1 → 2)-d-glucosidic linkages. It is concluded that the branches at O-2 are mainly single d-glucose units, whereas those occurring at O-3 may be longer than two glucose units, forming a highly branched structure having an average repeating- unit of 5 sugar residues.     

Structure of a new galactomannan from the ascocarps of Cordyceps cicadae shing

A water-soluble galactomannan (C-3), [α]_D²⁰ +30°, isolated from the rod-like ascocarps of Cordyceps cicadae, was determined to be homogeneous, and the molecular weight was estimated by gel filtration to be 27,000. The polysaccharide is composed of d-mannose and d-galactose in the molar ratio of 4:3. The results of methylation analysis, Smith degradation, stepwise hydrolysis with acid, and ¹³C-n.m.r. spectroscopy indicated that the polysaccharide is of highly branched structure, and composed of α-d-(1→2)-linked and α-d-(1→6)-linked mannopyranosyl residues in the core; some of these residues are substituted at O-6 and O-2 with terminal β-d-galactofuranosyl and α-d-mannopyranosyl groups, and with short chains of β-d-(1→2)-linked d-galactofuranosyl units.     

Structure of l-arabino-d-galactan-containing glycoproteins from radish leaves

Two l-arabino-d-galactan-containing glycoproteins having a potent inhibitory activity against eel anti-H agglutinin were isolated from the hot saline extracts of mature radish leaves and characterized to have a similar monosaccharide composition that consists of l-arabinose, d-galactose, l-fucose, 4-O-methyl-d-glucuronic acid, and d-glucuronic acid residues. The chemical structure features of the carbohydrate components were investigated by carboxyl group reduction, methylation, periodate oxidation, partial acid hydrolysis, and digestion with exo- and endo-glycosidases, which indicated a backbone chain of (1→3)-linked β-d-galactosyl residues, to which side chains consisting of α-(1→6)-linked d-galactosyl residues were attached. The α-l-arabinofuranosyl residues were attached as single nonreducing groups and as O-2- or O-3-linked residues to O-3 of the β-d-galactosyl residues of the side chains. Single α-l-fucopyranosyl end groups were linked to O-2 of the l-arabinofuranosyl residues, and the 4-O-methyl-β-d-glucopyranosyluronic acid end groups were linked to d-galactosyl residues. The O-α-l-fucopyranosyl-(1→2)-α-l-arabinofuranosyl end-groups were shown to be responsible for the serological, H-like activity of the l-arabino-d-galactan glycoproteins. Reductive alkaline degradation of the glycoconjugates showed that a large proportion of the polysaccharide chains is conjugated with the polypeptide backbone through a 3-O-d-galactosylserine linkage.     

Structural studies of the Rhizobium trifolii extracellular polysaccharide

The structure of the extracellular polysaccharide of Rhizobium trifolii has been investigated. Methylation analysis, sequential degradations by oxidation and elimination of oxidized residues, uronic acid degradation, and degradation by oxidation of the acetylated polysaccharide with chromium trioxide in acetic acid were the main methods used. It is proposed that the polysaccharide is composed of heptasaccharide repeating-units having the following structure:

An unusual feature is that some of the repeating units are incomplete and lack the terminal β-d-galactopyranosyl group. The polysaccharide contains O-acetyl groups (somewhat more than 1 mol. per unit), linked to O-2 and O-3 of 4-O-substituted d-glucopyranosyl chain-residues. A previous finding that the polysaccharide contains 2-deoxy-d-arabino-hexose (2-deoxy-d-glucose) residues is erroneous.     

Structural studies of an acidic polysaccharide from the mucin secreted by Drosera capensis

The polysaccharide of the mucin secreted by the leaves of Drosera capensis is composed of l-arabinose, d-xylose, d-galactose, d-mannose, and d-glucuronic acid in the molar ratio of 3.6:1.0:4.9:8.4:8.2. For structural elucidation, methylation analysis using g.l.c. and g.l.c.-m.s. was performed on the native, the carboxyl-reduced, and the degraded polysaccharides. Partial hydrolysis, periodate oxidation, chromium trioxide oxidation, and uronic acid degradation were also performed on the native and carboxyl-reduced polysaccharides. Partial hydrolysis of the native and carboxyl-reduced polysaccharides gave various oligosaccharides that were characterized and suggest a structure containing a d-glucurono-d-mannan backbone having a repeating unit → 4)-β-d-GlcpA-(1 → 2)-α-d-Manp-(1 →. l-Arabinose and d-xylose are present as nonreducing furanosyl and pyranosyl end-groups, respectively, both attached to O-3 of d-glucuronic acid residues of the backbone. d-Galactose is present as non-reducing pyranosyl end-group linked to O-3 of d-mannose residues.     

Structure of an l-arabino-d-xylan from the bark of Cinnamomum zeylanicum

An arabinoxylan isolated from the bark of Cinnamomum zeylanicum was composed of l-arabinose and d-xylose in the molar ratio 1.6:1.0. Partial hydrolysis furnished oligosaccharides which were characterised as α-d-Xylp-(1→3)-d-Ara, β-dXylp-(1→4)-d-Xyl, β-d-Xylp-(1→4)-β-d-Xylp-(1→4)-d-Xyl, β-d-Xylp-(1→4)-β-d-Xylp-(1→4)-β-d-Xylp-Xylp-(1→4)-d-Xyl, xylopentaose, and xylohexaose. Mild acid hydrolysis of the arabinoxylan gave a degraded polysaccharide consisting of l-arabinose (8%) and d-xyolse (92%). Methylation analysis indicated the degraded polysaccharide to be a linear (1→4)-linked d-xlan in which some xylopyranosyl residues were substituted at O-2 or O-3 with l-arabinofuranosyl groups. These data together with the results of methylation analysis and periodate oxidation of the arabinoxylan suggested that it contained a (1→4)-linked β-d-xylan backbone in which each xylopyranosyl residue was substituted both at O-2 and O-3 with l-arabinofuranosyl, 3-O-α-d-xylopyranosyl-l-arabinofuranosyl, and 3-O-l-arabinofuranosyl-l-arabinofuranosyl groups.     

Structural studies of water-soluble glycoproteins from Cannabis sativa L

Two carbohydrate-protein fractions, isolated from Cannabis sativa L. by extraction with water and chromatography on DEAE-cellulose, contained arabinose, galactose, glucose, mannose, galacturonic acid, 2-acetamido-2-deoxyglucose, and 2-acetamido-2-deoxygalactose. The structure of the carbohydrate moieties was investigated by methylation analysis and Smith degradation. A high percentage of end-groups indicates a large degree of branching, glucose and galactose being the main branch-points, linked at C-3 and C-6. The hexoses are also present as unbranched residues in the chain, largely as (1→3)- and (1→4)-linked units and as end-groups. Arabinofuranosyl units constitute the main part of the non-reducing end-groups, and are also present as part of the chain. The polysaccharide chains are probably linked to protein through the hydroxyl group of hydroxyproline.     

Smith degradation of gum exudates from some prosopis species

The polysaccharide of P. hymantophora has been shown to be composed of (1→4)-linked galactopyranosyl, (1→3)-linked galactopyranosyl, (1→3)-linked galactopyranosyl 2- and 4-sulphate and 2,6-disulphate residues. The (1→3)- and (1→4)-linked units are present in approximately equal amounts. The polysaccharide of P. hieroglyphica has been shown to possess (1→4)-linked galactopyranosyl, (1→3)-linked galactopyranosyl, and (1→3)-linked galactopyranosyl 2- and 4-sulphate residues. The (1→3)- and (1→4)-linked units are present in a 4:1 ratio. Both polysaccharides contain small proportions of non-reducing xylosyl end-groups.     

Methylation studies of the polysaccharides resulting from sequential Smith-degradations of the galactan from the snail strophocheilus oblongus

When the galactan from the albumen glands of the snail Strophocheilus oblongus was submitted to three Smith-degradations, the degraded polysaccharide, isolated in 6% yield, was much more linear. Methylation analysis showed that the Smith-degraded polysaccharide gave an increased percentage of 2,4,6-tri-, decreased percentages of 2,3,4,6-tetra- and 2,4-di-, and a large variation in the amount of 2,3,4-tri-O-methyl-d-galactose. The sugars in the polysaccharide which result in the formation of 2,3,4,6-tetra- and 2,3,4-tri-O-methyl-d-galactose are destroyed in subsequent degradation procedures. The above observations suggest that the degradation by periodate oxidation proceeds via non-reducing end-groups and through some internal residues that are exposed as the degradation proceeds. As a result of the overall process, new non-reducing end-groups are formed and new (1 → 6)-linked d-galactose residues are then exposed. The isolation of glycosides of low molecular weight supports the suggestion that the molecule, in the main, is sequentially degraded from the external layers.     

Structural study of the extracellular polysaccharide gum from Rhizobium strain CB744

The structure of the extracellular polysaccharide gum from nitrogen-fixing Rhizobium sp. strain CB744 (a member of the slow-growing Cowpea group) has been investigated. Gas-chromatographic analysis of the alditol acetates of the acid hydrolysate showed the gum to be composed of galactose, 4-O-methylgalactose, mannose, and glucose in the molar ratio of 1:2.5:3.5:7.0. The polysaccharide is unusual in that it contains no carbonyl substituent, although such substituents are common amongst polysaccharides produced by the slow-growing group. The native and de-branched polysaccharides were examined by methylation analysis. The anomeric configurations were determined by ¹³C-n.m.r. and oxidation by chromium trioxide. It is concluded that there are two β-(1→4)-linked glycopyranosyl residues for each α-(1→4)-linked mannopyranosyl residue, and that each mannose is substituted at O-6 by a β-galactopyranosyl residue, with 71% of the galactose groups being present as 4-O-methylgalactose.     

The preparation of a crystalline guanosine trialcohol and its sodium salt

A polysaccharide has been extracted from Cassia corymbosa seeds with cold, acidulated water, and purified to give a water-soluble product containing d-galactose and d-mannose in 4:7 molar ratio. Acid-catalyzed fragmentation, periodate oxidation, methylation, and enzymic hydrolysis showed that the seed gum has a branched structure consisting of a linear chain of β-d-(1→4)-linked mannopyranosyl units, some of which are substituted at O-6 by two α-d-(1→6) galactopyranosyl units mutually linked glycosidically. Methylation analysis of the galactomannan afforded 2,3,4-tri- and 2,3,4,6-tetra-O-methylgalactose, along with 2,3-di- and 2,3,6-tri-O-methylmannose, in the molar ratios of 2:2:2:5. Both the methylation and the periodate-oxidation studies showed ～36.4% of end groups. The significance of these results, together with the findings of partial hydrolysis with acid, are discussed, in relation to ascertaining the structure of the repeating unit of the polysaccharide.     

Structure of the d-galacto-d-mannan isolated from the seeds of Melilotus indica All

A d-galacto-d-mannan ([α]_D +72.0 and d-galactose-to-d-mannose ratio 1:1.14) was isolated from the seeds of Melilotus indica All., syn. M. parviflora Desf. The ¹H- and ¹³C-n.m.r., and i.r. spectra indicated the presence of α-d-galactopyranosyl and β-d-mannopyranosyl residues. Methylation of the polysaccharide, followed by hydrolysis, afforded, 2,3,4,6-tetra-, 2,3,6-tri-, 2,3-di-, and 3,4-di-O-methyl-d-mannose, and 2,3,4,6-tetra- and 2,3,6-tri-O-methyl-d-galactose in the molar ratios of 1:2:22:6:27:3. Periodate oxidation of the polysaccharide, followed by reduction and hydrolysis, gave erythritol (1 mol) and glycerol (1.24 mol). Partial acid hydrolysis of the polysaccharide afforded O-β-d-mannopyranosyl-(1→2)-d-mannopyranose, O-β-d-mannopyranosyl-(1→4)-d-mannopyranose, O-α-d-galactopyranosyl-(1→6)-d-mannopyranose, O-α-d-galactopyranosyl-(1→4)-d-galactopyranose, and O-α-d-galactopyranosyl-(1→6)-O-β-d-mannopyranosyl-(1→4)-d-mannopyranose. A highly branched structure having a mannan backbone composed of 36% of (1→4)- and 10% of (1→2)-linked β-d-mannopyranosyl units is proposed for the galactomannan.     

Bis(2-acetamido-3,4,6-tri-O-acetyl-2-deoxy-β-D-glucopyranosyl)amine obtained by a fusion reaction

A unique, alkali-soluble polysaccharide has been isolated from the cell walls of the basidiomycete Coprinus macrorhizus microsporus. The polysaccharide, which is primarily a glucan, contains a large proportion of α-(1→4)-linked d-glucose residues and a smaller amount of β-(1→3) and (1→6) linkages, as suggested by methylation, partial acid hydrolysis, periodate oxidation, and enzymic studies. Hydrolysis of the methylated polysaccharide gave equimolar amounts of 2,4-di- and 2,3-di-O-methyl-d-glucose; no 2,6-di-O-methyl-d-glucose was identified, indicating the absence of branch points joined through O-1, O-3, and O-4. The isolation and identification of 2-O-α- glucopyranosylerythritol from the periodate-oxidized polysaccharide suggests that segments of the a-(1→4)-linked d-glucose residues are joined by single (1→3)-linkages. An extracellular enzyme-preparation from Sporotrichum dimorphosporum (QM 806) containing both β-(1→3)- and α-(1→4)-d-glucanohydrolase activity released 76% of the reducing groups from the polysaccharide. The polysaccharide also contains minor proportions of xylose, mannose, 2-amino-2-deoxyglucose, and amino acids.     

Structure of the extracellular polysaccharide produced by the bacterium Alcaligenes (ATCC 31555) species

The extracellular anionic polysaccharide produced by the bacterium Alcaligenes (ATCC 31555) contains l-mannose, l-rhamnose, d-glucose, and d-glucuronic acid in the molar ratios 1.0:4.5:3.1:2.3. Analysis of the methylated and methylated, carboxyl-reduced polysaccharide indicated terminal non-reducing rhamnose and mannose, (1→4)-linked rhamnose, (1→3)- and (1→3,1→4)-linked glucose, and (1→4)-linked glucuronic acid to be present in the ratios 1.0:0.8:2.1:2.2:2.0:2.2. Partial acid hydrolysis and base-catalysed β-elimination gave a series of oligosaccharides that were isolated as their alkylated alditol derivatives by reverse-phase h.p.l.c. and characterised by f.a.b.-m.s., e.i.-m.s., and ¹H-n.m.r. spectroscopy. The repeating unit 1, excluding O-acyl groups, is proposed.

     

Structural investigations of the extracellular polysaccharide elaborated by s19, a Xanthomonas-type bacterium

The extracellular, acidic heteropolysaccharide from Xanthomonas S19 consists of D-glucuronic acid, D-glucose, D-galactose, and D-mannose residues in the approximate molar ratios of 1.6:3:1:1, plus acetyl groups liked to C-2 and/or C-3 of a large proportion of the glucose residues. Methylation studies showed that the glucose is present as non-reducing end-group also as 1,2- and 1,4-linked units, the galactose residues are solely 1,3-linked, a major proportion of the mannose residues are 1,2,4-linked and the rest 1,2-linked. A high proportion of the glucuronic acid units are 1,4-linked. Periodate oxidation confirmed the presence of these linkages. The disaccharides D-Glc-(1→4)-D-Glc,D-Glc-(1→2)-D-Man, D-Glc-(1→3)-D-Gal, D-Gal-(1→2)-D-Glc, D-GlcA-(1→4)-D-GlcA, and β-D-GlcA-(1→4)-D-Man were isolated from a partial hydrolysate of the polysaccharide, and characterised. The similarities and differences between this polysaccharide and those from other Xanthomonas species are discussed.     

The complete structure of the trifoliin a lectin-binding capsular polysaccharide of Rhizobium trifolii 843

The complete structure of the acidic, extracellular, capsular polysaccharide of Rhizobium trifolii 843 has been elucidated by a combination of chemical, enzymic, and spectroscopic methods, confirming an earlier proposed sugar sequence and assigning the locations of the acyl substituents. The polysaccharide was depolymerized by a lyase into octasaccharide units which were uniform in carbohydrate composition and linkage. These units also contained a uniform distribution of acetyl and pyruvic acetal [O-(1-carboxyethylidene)] groups, and half of them were further acylated with d-3-hydroxybutanoyl groups. A much smaller proportion (<5%) of the oligomers was further acylated by a second d-3-hydroxy-butanoyl group. The locations of the substituents were determined chemically and by J-correlated, ¹H-n.m.r. spectroscopy, proton nuclear Overhauser effect (n.O.e.)_ measurements, doubie-resonance ¹H-n.m.r. spectroscopy, and ¹³C-n.m.r. spectroscopy. The composition and structure of the carbohydrate chain were determined by methylation analysis using g.l.c.-m.s. fast-atom-bombardment mass spectrometry, and n.m.r. studies on the reduced, deacylated oligomer. Structural studies were supplemented by n.m.r. analyses on the original polymer. The oligosaccharides were found to be branched octasaccharides with four sugar residues in each branch, and the carbohydrate sequence agreed well with that expected from earlier work. In the abbreviated sequence and structure (1a), the sugar residues are labelled “a” through “h”. The main chain (a–d) is composed of a 4-deoxy-α-l-threo-hex-4-enopyranosyluronic acid group (a) that is linked to O-4 of a 3-O-acetyl-d-glucosyluronic acid residue (b) which is β-linked to O-4 of a d-glucosyl residue (c). Residue c is β-linked to O-4 of the branching d-linked to O-4 of a d-glucosyl residue (d). The side chain consists of a substituted d-galactosyl group (h) which is β-linked to O-3 of residue 9 of a β-(1→4)-linked d-glucose trisaccharide (fragment e–f–g). The reducing end of the resulting tetrasaccharide (e–f–g–h) is β-linked to O-6 of the branching d-glucose residue (d). In the native polymer, this branching residue is α-linked to O-4 of the modified d-glucuronic acid residue (a) which is the unsaturated sugar in the oligomer. A small proportion of the O-2 atoms of the acetylated d-glucosyluronic acid residues is acetylated because of ester migration. The two terminal sugars (g and h) of the branch chain bear 4,6-O-(1-carboxyethylidene) groups. The d-galactosyl groups of half of the oligomers are acylated by d-3-hydroxybutanoyl groups at O-3. About 5% of the oligomers bear a second d-3-hydroxybutanoyl group at O-2 of the d-galactosyl group (h).     

An acidic polysaccharide from the seeds of Ocimum adscendens

The mucilaginous polysaccharide-complex found in the seeds O. adscendens contains an acidic polysaccharide which has been isolated. It is composed of D-galactose (～ 20%), D-galacturonic acid (～ 35%) and L-rhamnose (～ 39%). Methylation analysis using GC and GC/MS, Smith degradation, isolation of an acidic oligosaccharide from partial hydrolysates, and enzymic hydrolysis using β-D-galactosidase indicated the polysaccharide backbone to be → 4)-GalpA-(1 → 2)-L-rhap-(1 →. Nearly two-thirds of the rhamnopyranosyl units are 0-4 substituted by D-galactopyranosyl non-reducing end groups.     

Location and identity of the acyl substituents on the extracellular polysaccharides of Rhizobium trifolii and Rhizobium leguminosarum

The basic structures of the extracellular polysaccharides of Rhizobium leguminosarum and Rhizobium trifolii were found to be identical, but their acylation patterns differ. Liquid hydrogen fluoride at −40° degrades the two polysaccharides to a series of oligosaccharides representing the repeating units of the polysaccharides and their higher homologs. At −23°, it degrades the polymers to a mixture of oligosaccharides from which a tetrasaccharide constituting a unit of the backbone of the polysaccharide, and a trisaccharide representing all but the non-reducing terminus of the side chain, could be readily purified. The location and identity of the acyl substituents were determined by ¹H-n.m.r. spectroscopy, methylation analysis, and f.a.b. mass spectrometry. The unusual substituent d-3-hydroxybutanoate was found esterified to O-3 of a terminal 4,6-O-pyruvic acetalated d-galactose in both strains of R. leguminosarum, and in one of the three strains of R. trifolii tested. All of the strains tested contained a 3-O-acetyl substituent on the (1→4)-β-d-glucopyranosyl residues in the backbone of the polysaccharide. Only the R. leguminosarum polysaccharides contained a combination of 2- and 3-O-acetyl groups on the branching sugar of the backbone of the polymer.     

Pyruvic acid-containing mono- and oligo-saccharides from rhizobium trifolii bart A

After partial, acid hydrolysis of the extracellular, acid polysaccharide from Rh. trifolii Bart A, the following products were isolated and characterized: 3,4-O-(1-carboxyethylidene)-d-galactose, 4,6-O-(1-carboxyethylidene)-d-galactose, 3-O-[3,4-O-(1-carboxyethylidene)-β-d)-galactopyranosyl]-d-glucose, 3-O-[4,6-O-(1-carboxyethylidene)-β-d-galactopyranosyl]-d-glucose, O-[3,4-O-(1-carboxyethylidene)-β-d-galactopyranosyl ]-(1→3)-O-d-glucopyranosyl-(1→4)-d-glucose, and O-[4,6-O-(1- carboxyethylidene)-β-d-galactopyranosyl]-(1→3)-O-β-d-glucopyranosyl-(1→4)-d-glucose. The presence of pyruvic acid linked either to O-3 and O-4 or to O-4 and O-6 of the d-galactopyranosyl group of these saccharides indicates that both structures may be present in the original polysaccharide.