Formation of Fungal Protoplast by the Reaction of Cell Wall Lytic Enzymes

Osmotically sensitive fungal protoplasts were liberated from the mycelium of several kinds of molds by mixed enzyme system of cellulase from Trichoderma viride and chitinase from the culture filtrate of Streptomyces sp. 115-5. Relatively higher number of protoplast were released from young mycelium of Zygomycetes strains than Ascomycetes strains by using 10 mM phosphate buffer (pH 6.0) and 0.6 M NaCl as osmotic stabilizer. Protoplasts were released through ruptures in the wall, initially at the apices, but later also from old party of the hyphae.