Summary: The formation of protoplasts from , by means of a lytic enzyme complex of , was considerably stimulated by adding thiols, including dithiothreitol, 2-mercaptoethanol and cysteamine, to the incubation medium. Treatment of the mycelium with these compounds before incubation with lytic enzymealso enhanced protoplast formation, dithiothreitol giving the most rapid release of protoplasts. The same stimulating effect could be obtained with a proteolytic enzyme. These results suggest the presence of proteinaceous material in the outer layers of the hyphal wall. protoplasts regenerated in liquid as well as solid media if a suitable osmotic stabilizer was present. Up to 20% of the protoplasts could grow into new mycelium.


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