Relative Abundance of Oligosaccharides in CandidaSpecies as Determined by Fluorophore-Assisted Carbohydrate Electrophoresis

  1. Jim E. Cutler
  1. Department of Microbiology, Montana State University, Bozeman, Montana 59817
  1. Fig. 1.
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    Fig. 1.

    Band fluorescence intensity as a function of carbohydrate concentration and oligomannoside length. (A) The relationship between band intensity and carbohydrate concentration was determined. Band fluorescence intensities of serial dilutions of maltose and maltotetraose were calculated and related to carbohydrate concentration for triplicate samples (r2 = 0.96140). (B) Nonpreferential ANTS labeling of maltooligosaccharides of various lengths was demonstrated. Aliquots of the maltooligosaccharide were derivatized under identical conditions for 0.5 to 12 h. The oligosaccharide concentration of the maltotetraose (G4) through maltonanose (G9) demonstrated an ANTS-labeling rate independent of oligomer length.

  2. Fig. 2.
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    Fig. 2.

    FACE results for the ANTS-labeled standard oligosaccharides and definition of the RMI. The RMI of monomeric glucose and the oligosaccharides derived from dextran were assigned unit values based on the number of hexose residues, i.e., glucose RMI 1.00, maltose RMI 2.00, etc. Every other migration position is expressed as a fraction of the linear distance between two adjacent reference points. Migration positions between the ANTS front, RMI 0.00, and glucose, RMI 1.00, were calculated in the same manner. The RMI of monomeric glucose (1.00), mannose (1.18), galactose (1.18), and arabinose (0.75) and of the α-1,4-oligoglucosides (⋄) and maltooligosaccharide (⌖) are indicated.

  3. Fig. 3.
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    Fig. 3.

    Relative abundances of oligosaccharides obtained by acid hydrolysis and β-elimination of Fehling-precipitated mannan fromC. albicans A9. The RMI values and the relative abundances of the individual moieties are indicated. Relative abundance was calculated using the linear regression (r2 = 0.96140) derived from known concentrations of maltotetraose (Fig. 1B).

  4. Fig. 4.
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    Fig. 4.

    FACE profile of oligosaccharides released by whole-cell acid hydrolysis of 16 Candida strains. The oligosaccharide banding patterns were specific for each Candida species. The RMI of selected bands illustrating band intensity differences, RMI 2.26 and RMI 3.10 (⌖) in C. stellatoidea, or band occurrence differences, RMI 1.82 (⋄) in C. glabrata 2001, are indicated. The RMI and relative abundances for all major bands (bands with intensities 10% greater than background) were calculated and tabulated (Table 1).

  5. Fig. 5.
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    Fig. 5.

    FACE profile of oligosaccharides released by whole-cell β-elimination of 16 Candida strains. The oligosaccharide banding patterns were specific for each Candida species. The RMI of bands that are specific for C. albicans 3153A andC. tropicalis (⋄) or more prominent in C. albicans A9 and C. stellatoidea (⌖) are indicated. The RMI for all major bands (bands with intensities 10% greater than background) were calculated and tabulated (Table 2).

  6. Fig. 6.
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    Fig. 6.

    FACE profile of oligosaccharides released by whole-cell acid hydrolysis and β-elimination of C. albicans A9 and four cell wall mutants, C. albicansA9-V2, C. albicans A9-V4, C. albicans A9-V8, andC. albicans A9-V10. The RMI of the fractionated products are indicated.

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  1. J. Clin. Microbiol. vol. 38 no. 8 2862-2869
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