SUMMARY: Osmotically fragile protoplasts have been prepared by the action of lytic enzymes on the mycelium of and The yield of protoplasts, based on DNA content, was up to 18 %. Pretreatment of the mycelium with a thiol compound was necessary with Cephalosporium but not with Penicillium. Electron micrographs indicated that the protoplasts contained all the intracellular organelles of the mycelium and were bounded only by a cytoplasmic membrane. The metabolic activity of the protoplasts, as measured by their respiration, ability to maintain intracellular amino-acid pools, and antibiotic production, was similar to that of control mycelium. -Valine and -α-aminoadipic acid, which are precursors of penicillin N and cephalosporin C, were taken up and concentrated by the protoplasts of , although the latter transported -valine less rapidly than did the corresponding mycelium. Protoplasts of took up -valine but not -α-aminoadipic acid or its δ-ester. There was little or no transport of the corresponding -amino acids by protoplasts of either organism.


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