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A simple method for quantitative determination of polysaccharides in fungal cell walls

Nature Protocols volume 1, pages 2995–3000 (2006)Cite this article

Abstract

A simple and reliable method for quantitative determination of cell wall polymers in fungal cell with an s.e.m. of 5% is described. This protocol is based on the hydrolysis by sulfuric acid of β-glucan, mannan, galactomannan and chitin present at different levels in the wall of yeasts and filamentous fungi into their corresponding monomers glucose, mannose, galactose and glucosamine. The released monosaccharides are subsequently separated and quantified by high-performance ionic chromatography coupled to pulse amperometry detection, with a detection limit of 1.0 μg ml−1. This procedure is well suited to screening a large collection of yeast mutants or to evaluating effects of environmental conditions on cell wall polysaccharide content. This procedure is also applicable to other fungal species, including Schizosaccharomyces pombe, Candida albicans and Aspergillus fumigatus. Results can be obtained in 3 d.

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Acknowledgements

This work was supported in part by grants from Fonds de Recherche Hoechst Marion Roussel (no. FRHMR2/9922) and the European Union (grants BIO4-CT95-0080 and QKL3-2000-01537). The author thanks Dr. Thierry Fontaine (Pasteur Institute, France) for his kind gift of purified cell wall samples from A. fumigatus and is grateful to Marie-Odile Loret for her expertise and help with HPIC measurement.

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  1. Laboratoire de Biotechnologie & Bioprocédés, UMR—CNRS 5504 and INRA 792, Institut National des Sciences Appliquées, 135 Avenue de Rangueil, Toulouse, cedex 04, 31077, France

    Jean Marie François

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Correspondence to Jean Marie François.

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François, J. A simple method for quantitative determination of polysaccharides in fungal cell walls. Nat Protoc 1, 2995–3000 (2006). https://doi.org/10.1038/nprot.2006.457

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