A simple method for quantitative determination of polysaccharides in fungal cell walls
A simple method for quantitative determination of polysaccharides in fungal cell walls
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DOI：
10.1038/nprot.2006.457
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Francois, Jean Marie
中科院分区：
文献类型：
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作者：
Francois, Jean Marie
用扫描电镜定量测定真菌细胞壁聚合物的方法。5%的描述。该方案基于硫酸将酵母和丝状真菌壁中不同水平存在的β-葡聚糖、甘露聚糖、半乳甘露聚糖和几丁质水解成其相应的单体葡萄糖、甘露糖、半乳糖和葡糖胺。释放的单糖，随后分离和定量的高性能离子色谱法耦合脉冲安培检测，检测限为1.0 μ g ml（-1）。该方法非常适合于筛选大量酵母突变体或评估环境条件对细胞壁多糖含量的影响。该程序也适用于其他真菌菌种，包括粟酒裂殖酵母、白色念珠菌和烟曲霉。结果可在3d内获得。
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 b-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 mu g 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.