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Structural analysis of the exopolysaccharide produced by Streptococcus thermophilus ST1 solely by NMR spectroscopy

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Abstract

The use of lactic acid bacteria in fermentation of milk results in favorable physical and rheological properties due to in situ exopolysaccharide (EPS) production. The EPS from S. thermophilus ST1 produces highly viscous aqueous solutions and its structure has been investigated by NMR spectroscopy. Notably, all aspects of the elucidation of its primary structure including component analysis and absolute configuration of the constituent monosaccharides were carried out by NMR spectroscopy. An array of techniques was utilized including, inter alia, PANSY and NOESY-HSQC TILT experiments. The EPS is composed of hexasaccharide repeating units with the following structure: → 3)[α-d-Glcp-(1 → 4)]-β-d-Galp-(1 → 4)-β-d-Glcp-(1 → 4)[β-d-Galf-(1 → 6)]-β-d-Glcp-(1 → 6)-β-d-Glcp-(1 →, in which the residues in square brackets are terminal groups substituting backbone sugar residues that consequently are branch-points in the repeating unit of the polymer. Thus, the EPS consists of a backbone of four sugar residues with two terminal sugar residues making up two side-chains of the repeating unit. The molecular mass of the polymer was determined using translational diffusion experiments which resulted in Mw = 62 kDa, corresponding to 64 repeating units in the EPS.

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Abbreviations

EPS:

Exopolysaccharide

TILT:

Time-domain increments linked together

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Acknowledgments

This work was supported by grants from the Swedish Research Council (VR), The Knut and Alice Wallenberg Foundation, Carl Tryggers Stiftelse för Vetenskaplig Forskning, Magn. Bergvalls Stiftelse, the Academy of Finland (Project number 210653) and the Sino-Finnish Cooperative Project (2006DFA32620). The Swedish NMR Centre at Göteborg University is thanked for access to a high-field spectrometer. We thank Dr. Jesper Lind for assistance with the DLS measurements.

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Authors and Affiliations

  1. Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, 106 91, Stockholm, Sweden

    Elin Säwén & Göran Widmalm

  2. Department of Food Technology, University of Helsinki, 00014, Helsinki, Finland

    Eine Huttunen & Xue Zhang

  3. Center of Agro-food Technology, Northeast Agricultural Research Center of China, 130124, Changchun, Jilin, People’s Republic of China

    Zhennai Yang

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  1. Elin Säwén
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  2. Eine Huttunen
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  3. Xue Zhang
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  4. Zhennai Yang
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  5. Göran Widmalm
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Correspondence to Göran Widmalm.

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Säwén, E., Huttunen, E., Zhang, X. et al. Structural analysis of the exopolysaccharide produced by Streptococcus thermophilus ST1 solely by NMR spectroscopy. J Biomol NMR 47, 125–134 (2010). https://doi.org/10.1007/s10858-010-9413-0

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