Treatment of fungal GlcCer in vitro with Cerezyme.

A) Enzymatic reaction leading to the degradation of GlcCer in Cn catalyzed by beta-glucosidase (Cerezyme). B) Extracted cryptococcal GlcCer (8 µg) was treated in vitro with 200 mU/ml of Cerezyme for 1 hour at 37°C. The reaction was stopped, lipids extracted, and loaded on a TLC with a soy standard for GlcCer. The GlcCer was visualized with a resorcinol spray. C) The same amount of GlcCer was treated with 200 mU/ml of Cerezyme and the amount of GlcCer was analyzed with mass spectrometry, along with the amount of the reaction product, α-OH-Δ8-Me-Ceramide (Me-Ceramide). The GlcCer treated with Cerezyme showed complete hydrolysis in these conditions.