Formation and regeneration of Penicillium chrysogenum protoplasts.

Abstract

1. High yields of protoplasts from Penicillium chrysogenum Wisc. 49,2105 were obtained by using a combined enzyme system containing either Cellulase from Oxoporus plus an extract of Helix pomatia gut juice, or cellulase plus an enzyme preparation from the culture filtrate of Streptomyces graminofaciens ATCC 12,705. When 15 h old mycelium was incubated with one of these enzyme systems in 0.55 M NaCl at pH 5.6 nearly all of the mycelium was transformed into protoplasts within 4–5 h. 2. The process of protoplast regeneration was studied both on solid and in liquid media. Approximately 50% of the protoplasts regenerated within 8–10 h. Addition of yeast extract to the medium accelerated the speed of regeneration. Microscopically, 3 patterns were seen by which protoplasts could regenerate to normal mycelium.