Abstract
Cell wall composition is a useful marker to indicate taxonomic and phylogenetic affiliations among fungi. Four main methods have been applied to analyze the carbohydrate composition of the yeast cell wall, which include gas chromatographic analysis of acid hydrolyzates of whole cells with derivatization using capillary columns; gas chromatographic analysis of acid hydrolyzates of purified cell walls with derivatization; high performance liquid chromatographic (HPLC) analysis of acid hydrolyzates of whole cells without derivatization; and high performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD) of cell wall neutral sugars without derivatization. In the first method, the cells are hydrolyzed with 1 N HCl for 12 hours at l00° C, during which monomers are formed. Neutral polysaccharides are hydrolyzed completely at low concentrations of HCl (1 N), whereas chitin is converted to glucosamine at high concentrations of HCl (5 N). After hydrolysis, the solubilized components are trimethylsilylated (TMS) prior to gas-liquid chromatography. In the second method, the cell walls are isolated and purified before further processing. The third method was developed by Suzuki and Nakase, which used HPLC analysis of whole-cell hydrolyzates without derivatization. In this method, whole cells are hydrolyzed with trifluoroacetic acid (TFA) and directly analyzed by using HPLC.
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