Abstract
A novel method for preparation ofNeurospora crassa protoplasts has been developed based on treatment of hyphae with an inducible enzyme system ofArthrobacter GJM-1 that had been obtained by growing the bacterium onNeurospora cells as the sole carbon source. The lytic system was characterized for enzyme activities relevant to hydrolysis of major cell-wall components ofN. crassa, such as 1,3-β-d-glucanase, α-mannanase, chitinase, proteinase, α-amylase and β-d-glucosidase. The optimal conditions for protoplast formation from young hyphae ofN. crassa (10 mg dry weight) were 180 min treatment by 40 mg/ml of freeze-dried lytic system. Protoplasts which were released from both hyphal tips and sides were found not to contain cell-wall material, as judged by electron microscopy and Calcofluor binding. When freed of the lytic system, 50% of protoplasts regenerated a mycelium.
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