Base hydrolysis of phosphodiester bonds in pneumococcal polysaccharides

Pujar, Narahari S.; Huang, Ngan F.; Daniels, Christopher L.; Dieter, Lance; Gayton, Marshall; Lee, Ann L.

doi:10.1002/bip.20087

Cited by 24 publications

(12 citation statements)

References 29 publications

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“…The walls of gram-positive bacteria may contain one or more secondary or accessory polymers that are attached via phosphodiester bonds to peptidoglycan (i.e., teichoic acid or teichuronic acids)22. Although similar phosphodiester bonds have been described for polysaccharides from pneumococci23 and meningococci24, to the best of our knowledge, no previous paper has described a phosphodiester bond within a polysaccharide of a Lactobacillus . L .…”

Section: Discussionmentioning

confidence: 95%

Structural and immunomodulatory differences among lactobacilli exopolysaccharides isolated from intestines of mice with experimentally induced inflammatory bowel disease

Górska

Sandström

Wojas-Turek

et al. 2016

Sci Rep

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Characteristic changes in the microbiota biostructure and a decreased tolerance to intestinal bacteria have been associated with inflammatory bowel disease (IBD). However, few studies have examined the constituents of the intestinal microbiota, including the surface molecules of the bacteria, in healthy and IBD subsets. Here, we compare the chemical structures and immunomodulatory properties of the exopolysaccharides (EPS) of lactobacilli isolated from mice with induced IBD (IBD “+”) versus those of healthy mice (IBD “−”). Classical structural analyses were performed using nuclear magnetic resonance spectroscopy and mass spectrometry. Immunomodulatory properties were assessed by stimulation of dendritic cells derived from mouse bone marrow or human peripheral mononuclear blood cells. Our results revealed that EPS produced by IBD “+” species are structurally different from those isolated from IBD “−”. Moreover, the structurally different EPS generate different immune responses by dendritic cells. We speculate that resident strains could, upon gut inflammation, switch to producing EPS with specific motifs that are absent from lactobacilli IBD “−”, and/or that bacteria with a particular EPS structure might inhabit the inflamed intestinal mucosa. This study may shed light on the role of EPS in IBD and help the development of a specific probiotic therapy for this disease.

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Section: Discussionmentioning

confidence: 95%

Structural and immunomodulatory differences among lactobacilli exopolysaccharides isolated from intestines of mice with experimentally induced inflammatory bowel disease

Górska

Sandström

Wojas-Turek

et al. 2016

Sci Rep

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“…The generated oligosaccharides are isolated and structurally analyzed by a variety of chemical and physical methods. This strategy has been extensively used by our group and others 26, 27, 38–42. To degrade polysaccharides, two common approaches can be employed, chemical and enzymatic.…”

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confidence: 99%

In‐source fragmentation and analysis of polysaccharides by capillary electrophoresis/mass spectrometry

Wang

Altman

2005

Rapid Comm Mass Spectrometry

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In order to develop a robust and easy-to-use technique for characterization of bacterial polysaccharides, a pseudo-hydrolysis strategy was investigated. Based on in-source collision-induced dissociation, polysaccharide molecular ions were fragmented within the orifice-skimmer region of an electrospray ionization (ESI) mass spectrometer. The fragment ions thus generated were then analyzed similarly to the conventional ESI mass spectrometry approach. MS/MS scanning was applied to obtain product-ion spectra of the primary fragments for sequencing. To further improve the sensitivity and separation of polysaccharides from other components in the samples, a pressure-assisted capillary electrophoresis/mass spectrometry (CE/MS) system was employed. Using bacterial polysaccharides as model compounds, the mass spectra obtained for polysaccharide repeating units generated through chemical hydrolysis and in-source fragmentation were directly compared, both in positive and negative ion modes. With the additional separation of impurities provided by CE, the success of this technique has been demonstrated for structural analysis of O-chain polysaccharides (O-PS) and capsular polysaccharides (CPS). In-source fragmentation was applied to promote the formation of structurally relevant repeating units of heterogeneous CPS that would remain undetected using conventional ESI conditions. This approach was proven to be particularly useful for probing the subtle structural differences in monosaccharide composition and functionalities arising across bacterial serotypes.

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“…As discussed in Sect. The effect of base hydrolysis on the molecular size of these phosphodiester-containing polysaccharides was studied by use of SEC-HPLC-RI/MALLS (Pujar et al 2004). The instability of polysaccharides is associated with a phosphodiester linkage and conditions for improving their stability (use of DMSO and sucrose) during reductive amination conjugation have been reported (Prasad 2011).…”

Section: Stability Testingmentioning

confidence: 99%