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Glycan structure of the S-layer glycoprotein ofBacillus sp. L420-91

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Abstract

Preliminary taxonomic characterization of isolate L420-91 has revealed that this organism is closely related to the speciesBacillus aneurinolyticus. The bacterium is covered by a squarely arranged crystalline surface layer composed of identical glycoprotein subunits with an apparent molecular mass in the range of 109 kDa. A total carbohydrate content of approximately 3.5% (wt/wt) was determined in the purified surface layer glycoprotein. Glycopeptides were obtained after exhaustive Pronase digestion and purification including gel filtration, ion exchange chromatography and HPLC. From the combined evidence of composition analysis, Smith degradation and nuclear magnetic resonance spectroscopy experiments we propose the following structure for the glycan chain of the surface layer glycoprotein:

$$\begin{array}{*{20}c} { \to 3) - \alpha - D - Rhap - (1 \to 3) - \alpha - D - Rhap - (1 \to 2) - \alpha - D - Rhap - (1 \to 2) - \alpha - D - Rhap - (1 \to } \\ {\begin{array}{*{20}c} {\begin{array}{*{20}c} 2 \\ \uparrow \\ 1 \\ \end{array} } & {\begin{array}{*{20}c} 2 \\ \uparrow \\ 1 \\ \end{array} } \\ \end{array} } \\ {\alpha - D - Fucp3NAc \alpha - D - Fucp3NAc} \\ \end{array}$$

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Abbreviations

S-layer:

crystalline surface layer

SDS-PAGE:

sodium dodecyl sulphate polyacrylamide gel electrophoresis

RP-HPLC:

reversed phase high performance liquid chromatography

HPAEC:

high performance anion exchange chromatography

TFA:

trifluoroacetic acid

NMR:

nuclear magnetic resonance spectroscopy

COSY:

correlated spectroscopy

TOCSY:

total correlated spectroscopy

NOE:

nuclear Overhauser enhancement

dRha:

d-rhamnose

dFuc3NAc:

3-acetamido-3-deoxy-d-fucose

Man:

mannose

Gro:

glycerol

Thr:

threonine

Gly:

glycine

Tyr:

tyrosine

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Authors and Affiliations

  1. Institut für Chemie, Universität für Bodenkultur, A-1180, Wien, Austria

    Paul Kosma

  2. Zentrum für Ultrastrukturforschung und Ludwig Boltzmann-Institut für Molekulare Nanotechnologie, Universität für Bodenkultur, A-1180, Wien, Austria

    Christina Neuninger & Paul Messner

  3. Scientific Software Comp., A-1140, Wien, Austria

    Rudolf Christian

  4. Sandoz Forschungsinstitut Wien, A-1235, Wien, Austria

    Gerhard Schulz

Authors
  1. Paul Kosma
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  2. Christina Neuninger
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  3. Rudolf Christian
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  4. Gerhard Schulz
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  5. Paul Messner
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Kosma, P., Neuninger, C., Christian, R. et al. Glycan structure of the S-layer glycoprotein ofBacillus sp. L420-91. Glycoconjugate J 12, 99–107 (1995). https://doi.org/10.1007/BF00731875

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  • DOI: https://doi.org/10.1007/BF00731875

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