What Is the Specific Protocol for the Extraction of Fungal Cell Wall Polysaccharides?
The extraction of fungal cell wall polysaccharides typically involves several sequential steps: cultivation and harvesting of fungal biomass, isolation of the cell wall, extraction of polysaccharides from the wall material, and purification of the extracted polysaccharides. Depending on the fungal species, additional specific treatments may be required to ensure efficient and complete recovery of polysaccharides. Here, we present a detailed protocol using a commonly studied fungus, Saccharomyces cerevisiae (yeast), as a representative example:
Cultivation and Harvesting of Yeast Cells
An appropriate S. cerevisiae strain is selected and cultured using either liquid or solid media under optimal temperature, pH, and incubation time. The yeast biomass is then harvested via centrifugation or other suitable methods.
Cell Wall Isolation
The collected yeast cells are centrifuged, and mechanical or enzymatic disruption methods—such as ultrasonication or enzymolysis—are applied to rupture the cells, allowing separation of the cell wall from intracellular components.
Extraction of Cell Wall Polysaccharides
The isolated cell wall material is subjected to hot water extraction using acidic or alkaline conditions, for instance, 2% acetic acid at elevated temperature for 2–3 hours. The extraction may be repeated multiple times to maximize polysaccharide yield.
Purification of Polysaccharides
Crude extracts are purified through commonly employed techniques such as ethanol precipitation, ion-exchange chromatography, and gel filtration chromatography. These approaches facilitate the removal of contaminants and yield purified polysaccharides.
Characterization and Analysis of Polysaccharides
The purified polysaccharides are analyzed using high-performance liquid chromatography (HPLC), gas chromatography (GC), nuclear magnetic resonance (NMR) spectroscopy, and other analytical techniques to determine their composition, structural features, and molecular weight.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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