Liberation of protoplasts from mycelium of Neurospora crassa by means of enzymes obtained from autolyzed cultures of this fungus

Abstract

Spherical protoplast-like structures were released from Neuropsora crassa mycelia under the influence of cell-wall-lytic enzymes obtained from autolyzed cultures of this fungus, in 50 mM borate-citrate-phosphate buffer, pH 5.5 with 0.5-0.8 M mannitol for appropriate osmotic pressure. The protoplasts retain their morphological aspect for several days at 4 C, or for a longer time when they are washed by centrifugation and incubated with 0.8 M mannitol. The free protoplasts are sensitive to osmotic shock and lyse immediately when placed in distilled water; the vacuoles and other cytoplasmatic material remain intact for some time.