Re-examination of cellular cyclic β-1,2-glucans of Rhizobiaceae: Distribution of ring sizes and degrees of glycerol-1-phosphate substitution

Zevenhuizen, L.P.T.M.; Veldhuizen, A. van; Fokkens, R.H.

doi:10.1007/bf00403952

Cited by 34 publications

(29 citation statements)

References 19 publications

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“…Resonances of low intensity at 63.7 and 67.7 p.p.m. could be assigned to the C3-and C,-carbons of the glycerol-phosphate substitutions being present on part of the C,-glucoses of the anionic cyclic glucans in R. meliloti (Zevenhuizen et al, 1990). The C,-resonance of the glycerol-phosphate substitution at 7 1.9 p.p.m.…”

Section: In Vivo '3c-nmrmentioning

confidence: 94%

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Response of intracellular carbohydrates to a NaCl shock in Rhizobium leguminosarum biovar trifolii TA-1 and Rhizobium meliloti SU-47

Breedveld

Dijkema

Zevenhuizen

et al. 1993

Journal of General Microbiology

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~ ~The dynamic response of cellular carbohydrates to a NaCl shock in Rhizobium leguminosarum biovar trifolii TA-1 (0.25 M-NaCI) and Rhizobium meliloti SU-47 (0.4 M-NaCl) grown in NaC1-free medium was investigated in nongrowing cell cultures and in cell suspensions, using in vivo NMR. After transferring cells grown in a NaC1-free medium to a glutamic-acid-free medium containing mannitol and NaCI, both strains immediately responded to the increased osmotic pressure by augmenting the cellular trehalose content of the cell. Without mannitol in the medium trehalose synthesis was slower, but clearly detectable. Its synthesis paralleled the breakdown of the reserve materials glycogen and poly-P-hydroxybutyric acid (PHB). NMR experiments with 25-fold-concentrated cell suspensions using '3C,-mannitol as substrate revealed that 15-20 % of the trehalose synthesized was derived from mannitol, but 8&85% was from other sources. Trehalose was mainly formed from the internal pool of glycogen and/or PHB, whether mannitol was present or not, and reached 135 and 280 pg (mg cell protein)-' in the strains TA-1 and SU-47, respectively. At low osmolarity, intracellular trehalose was metabolized by strains TA-1 and SU-47. Intracellularly accumulated phosphoglycerol-substituted and neutral cyclic (1,2)-/?-glucans of SU-47 cells grown in the absence of NaCl were neither degraded nor excreted after exposure to NaC1. Strain TA-1, which only makes neutral cyclic (1,2)-P-glucans, continued to synthesize and excrete cyclic (1,2)-P-glucans after exposure to NaCI. By using in vivo 31P-NMR, a sharp peak at 1-34 p.p.m. was present in cell suspensions of strain SU-47. This peak, representing glycerol-1-phosphate-substituted cyclic glucans, was absent in strain TA-1.

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Section: In Vivo '3c-nmrmentioning

confidence: 94%

“…The intracellular concentration of cyclic (1,2)-P-glucans in the cell suspension of strain SU-47 was 260 pg (mg cell protein)-' or 3mg (ml sus- (Fig. l a ) could be easily assigned (Zevenhuizen et al, 1990). Resonances of low intensity at 63.7 and 67.7 p.p.m.…”

Section: In Vivo '3c-nmrmentioning

confidence: 95%

Response of intracellular carbohydrates to a NaCl shock in Rhizobium leguminosarum biovar trifolii TA-1 and Rhizobium meliloti SU-47

Breedveld

Dijkema

Zevenhuizen

et al. 1993

Journal of General Microbiology

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“…The degree of substitution varies among species; for example, cyclic glucans produced by the broad-host-range Rhizobium sp. strain GRH2 and by certain strains of Rhizobium leguminosarum contain no anionic substituents (30,45). Bradyrhizobium japonicum cyclic ␤-1,6-␤-1,3-glucans are neutral but substituted with the zwitterionic phosphocholine (35).…”

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The Brucella abortus Cyclic β-1,2-Glucan Virulence Factor Is Substituted with O-Ester-Linked Succinyl Residues

et al. 2006

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Brucella periplasmic cyclic ␤-1,2-glucan plays an important role during bacterium-host interaction. Nuclear magnetic resonance spectrometry analysis, thin-layer chromatography, and DEAE-Sephadex chromatography were used to characterize Brucella abortus cyclic glucan. In the present study, we report that a fraction of B. abortus cyclic ␤-1,2-glucan is substituted with succinyl residues, which confer anionic character on the cyclic ␤-1,2-glucan. The oligosaccharide backbone is substituted at C-6 positions with an average of two succinyl residues per glucan molecule. This O-ester-linked succinyl residue is the only substituent of Brucella cyclic glucan. A B. abortus open reading frame (BAB1_1718) homologous to Rhodobacter sphaeroides glucan succinyltransferase (OpgC) was identified as the gene encoding the enzyme responsible for cyclic glucan modification. This gene was named cgm for cyclic glucan modifier and is highly conserved in Brucella melitensis and Brucella suis. Nucleotide sequencing revealed that B. abortus cgm consists of a 1,182-bp open reading frame coding for a predicted membrane protein of 393 amino acid residues (42.7 kDa) 39% identical to Rhodobacter sphaeroides succinyltransferase. cgm null mutants in B. abortus strains 2308 and S19 produced neutral glucans without succinyl residues, confirming the identity of this protein as the cyclic-glucan succinyltransferase enzyme. In this study, we demonstrate that succinyl substituents of cyclic ␤-1,2-glucan of B. abortus are necessary for hypo-osmotic adaptation. On the other hand, intracellular multiplication and mouse spleen colonization are not affected in cgm mutants, indicating that cyclic-␤-1,2-glucan succinylation is not required for virulence and suggesting that no lowosmotic stress conditions must be overcome during infection.Brucella spp. are gram-negative, facultative intracellular bacteria that cause a chronic zoonotic disease known worldwide as brucellosis. Brucella abortus, the etiological agent of bovine brucellosis, causes abortion and infertility in cattle and undulant fever in humans (17). The human disease is also caused by Brucella melitensis, Brucella suis, and Brucella canis. Brucella spp. are intracellular pathogens that invade and proliferate within host cells; virulence is associated with the ability to multiply inside professional and nonprofessional phagocytic cells (40).Cyclic glucans appear to be important as intrinsic components of gram-negative bacterial cell envelopes (7). Beyond their common features, glucans show an unexpected structural diversity (4). The role of cyclic glucan in gram-negative bacterial cell envelopes is still controversial. Cyclic-glucan mutants have a highly pleiotropic phenotype, suggesting an overall alteration of their cell envelope properties. Mutants deficient in cyclic-glucan production were found to be attenuated or avirulent compared to their wild-type parent strains, suggesting that the cyclic glucan is an important virulence attribute for the bacteria in their interaction with eukaryotic host...

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“…The EPSs produced by rhizobia are high-molecular-weight heteropolysaccharides composed of small repeating units that contain either uronic acids or noncarbohydrate substituents, such as succinate and pyruvate, as the acidic functions (4,22,29). The cyclic 3-glucans are commonly harbored within the periplasmic space but may also be secreted; consequently, they are found in both the growth media and the bacterial pellet of cultured cells (42). The role of the 13-glucans in symbiosis remains a moot subject.…”

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Rhizobium fredii and Rhizobium meliloti produce 3-deoxy-D-manno-2-octulosonic acid-containing polysaccharides that are structurally analogous to group II K antigens (capsular polysaccharides) found in Escherichia coli

1993

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The polysaccharide components from cultured cells of Rhizobium fredii USDA205 and Rhizobium meliloti AK631 were extracted with hot phenol-water and separated by repetitive gel filtration chromatography. Polyacrylamide gel electrophoresis, nuclear magnetic resonance spectrometry, and gas chromatography analyses showed that both of these bacterial species produce unique polysaccharides that contain a high proportion of 3-deoxy-D-manno-2-octulosonic acid (Kdo). These polysaccharides, which constituted a major portion of the extracted carbohydrate, are not excreted into the growth media (i.e., they are not extracellular polysaccharides) and are structurally distinct from the lipopolysaccharides. The primary structure of the preponderant polysaccharide from R. fredii USDA205 was determined by high-performance anion-exchange liquid chromatography, nuclear magnetic resonance spectrometry, fast atom bombardment-mass spectrometry, and gas chromatography-mass spectrometry; it consists of repeating units of [-- The gram-negative bacteria of the Rhizobiaceae family include the nitrogen-fixing symbionts of legumes (Rhizobium,Azorhizobium, and Bradyrhizobium spp.) and the pathogenic Agrobacterium genus. These species produce various types of extracellular polysaccharides (EPSs) and cell surface polysaccharides that appear to be important factors in symbiotic and pathogenic effectiveness.Lipopolysaccharides (LPSs), unique to gram-negative bacteria, are outer membrane inclusions that consist of (i) a glycolipid membrane anchor, which contains characteristic Il-hydroxy fatty acids on a glucosamine backbone (lipid A);(ii) a core oligosaccharide that is linked to the lipid A via 3-deoxy-D-manno-2-octulosonic acid (Kdo) and is highly conserved within a species; and (iii) a highly variable 0 antigen. Polyacrylamide gel electrophoresis (PAGE)-silver stain analyses of rhizobial LPSs frequently result in banding regions (5, 9), indicating that the 0 antigens of these LPSs may commonly consist of oligosaccharides of specific lengths.Rhizobia also produce acidic EPSs and cyclic P-glucans.EPSs are secreted compounds and are isolated from the growth media of cultured cells. The EPSs produced by rhizobia are high-molecular-weight heteropolysaccharides composed of small repeating units that contain either uronic acids or noncarbohydrate substituents, such as succinate and pyruvate, as the acidic functions (4, 22, 29). The cyclic 3-glucans are commonly harbored within the periplasmic space but may also be secreted; consequently, they are found in both the growth media and the bacterial pellet of cultured cells (42). The role of the 13-glucans in symbiosis remains a moot subject. * Corresponding author.Numerous studies have shown that bacterial LPS and EPS are important in the infection and nodulation processes; however, the symbiotic phenotype of LPS-and EPS-defective microsymbionts appears to depend on the type of nodule formed by the host.(i) Inoculation of determinate nodule-forming host plants with rhizobial LPS mutants that are affected ...

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Re-examination of cellular cyclic β-1,2-glucans of Rhizobiaceae: Distribution of ring sizes and degrees of glycerol-1-phosphate substitution

Cited by 34 publications

References 19 publications

Response of intracellular carbohydrates to a NaCl shock in Rhizobium leguminosarum biovar trifolii TA-1 and Rhizobium meliloti SU-47

Response of intracellular carbohydrates to a NaCl shock in Rhizobium leguminosarum biovar trifolii TA-1 and Rhizobium meliloti SU-47

The Brucella abortus Cyclic β-1,2-Glucan Virulence Factor Is Substituted with O-Ester-Linked Succinyl Residues

Rhizobium fredii and Rhizobium meliloti produce 3-deoxy-D-manno-2-octulosonic acid-containing polysaccharides that are structurally analogous to group II K antigens (capsular polysaccharides) found in Escherichia coli

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