Digestion of yeast and fungal cell walls is necessary for many experimental procedures including spheroplasting, immunofluorescence, transformation, protein purification, and others. The use of lytic enzymes like Zymolyase is routinely used for digestion. The Zymolyase from Zymo Research is prepared from Arthrobacter luteus, lyophilized, and packaged with a resuspension buffer. The buffer has been optimized to confer maximal levels of enzymatic activity. The main activities of the enzyme are β-1,3 glucanase and β-1,3-glucan laminaripentao-hydrolase, which hydrolyze glucose polymers at the β-1,3-glucan linkages releasing laminaripentaose as the principal product. Optimal Zymolyase activity is at 30° – 37°C; lytic activity ceases at higher temperatures. R-Zymolyase includes 0.5 U/µl RNase A when reconstituted. Susceptible fungal genera: Asbya, Candida, Debaryomyces, Eremothecium, Endomyces, Hansenula, Hanseniaspora, Kloekera, Kluyveromyces, Lipomyces, Metschikowia, Pichia, Pullularia, Saccharomyces, Saccharomyces, Saccharomycopsis, Schizosaccahromyces, and Torulopsis.