A simple method for quantitative determination of polysaccharides in fungal cell walls.
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 beta-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 mug 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.
François
Jean Marie
JM
Laboratoire de Biotechnologie & Bioprocédés, UMR-CNRS 5504 and INRA 792, Institut National des Sciences Appliquées, 135 Avenue de Rangueil, 31077 Toulouse cedex 04, France.
D002473:Cell Wall / Q000737:chemistry*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004560:Electricity; D005658:Fungi / Q000737:chemistry*; D011134:Polysaccharides / Q000032:analysis*
D011134:Polysaccharides