I am studying the enzyme trehalase and its effect on trehalose in mutant Crypotococcus fungal cells. Cryptococcus is a fungus that has a tendency to produce meningoencephalitis in immunocompromised hosts. Trehalose is a disaccharide that helps the fungus to survive by increasing its resistance to heat and preventing the denaturation of important proteins. Trehalase is the enzyme that breaks down trehalose into glucose.
In the lab, I have been growing mutant cells in YDP growth medium and comparing trehalose levels in the cells treated with trehalase with the cells treated without trehalase. I do this by preparing a glucose assay and measuring the glucose levels in each cell, which indirectly measures trehalose levels (because trehalase breaks down trehalose into glucose, higher glucose levels means less trehalose). Cells that show lower levels of trehalose in trehalase-treated cells versus non-trehalase-treated cells indicate that these cells have a mutation that affects how much trehalose is produced.