Extracting Mushroom Polysaccharides for Intended Benefits of Immunostimulation and Therapeutic Uses

The use of mushrooms adjuvant in clinical studies to boost immunity and prevent malignancies and viral infections has been promoted by the discovery of bioactive compounds with immunomodulatory capabilities. It is possible to genetically modify mushrooms to produce the desired bioactive compounds in large numbers, which may then be separated for use in both clinical and commercial settings. The purpose of this study is to identify a novel bioactive molecule in Calocybe Indica mushroom sample, which we found to have immunomodulatory properties and potential clinical uses. The research question in action is how to extract and identify polysaccharides and their composition from Calocybe Indica. The methodology used was a hot water extraction set up to extract water-soluble constituents, the concentration of extract, and precipitation of polysaccharides ethanol and centrifugation. Next, to identify the monosaccharide composition, three Thin Layer Chromatography methods were selected, and the Aniline Diphenylamine Phosphoric Acid method produced the best results when tested with sugar samples. Currently, the ongoing experiment involves hydrolyzing the polysaccharides into monomers and conducting a Thin-Layer Chromatography experiment with the hydrolyzed sample to determine the composition of the Calocybe Indica monomers. To further confirm the results of TLC (Thin Layer Chromatography), HPLC (High-Performance Liquid Chromatography) protocol has been developed and is currently being tested. The future work of this project involves optimization of TLC and HPLC protocols, followed by fractionation by gel-filtration chromatography and MALDI analysis of Calocybe Indica extracts.