Release of protoplasts from the galactose oxidase producing mold, Dactylium dendroides

Osmotically sensitive protoplasts were released from the mycelium of Dactylium dendroides, using Megalobolinus paranaguensis digestive juice as a lytic enzyme. The conditions for obtaining stable protoplasts were determined. The maximum number of protoplasts was obtained from 15-h growing mycelium, using MgSO4 as osmotic stabilizer, in the presence of 0.015 M Sorensen phosphate buffer, pH 5.6. MgSO4 proved to be of considerable importance in the release of protoplasts in this fungus. Regeneration of the protoplasts was demonstrated in normal growth medium supplemented with 0.8 M mannitol and 2.5% agar.