Abstract
A reproducible and reliable protoplasting and regeneration condition for the genus Actinomadura strains, which is extraordinarily resistant to lysozyme, has been studied. Protoplasts were prepared from all strains examined by use of both lysozyme and N-acetylmuramidase only when they were grown in the presence of sucrose (10∼30%) and glycine (0.3%). The optimum concentration of sucrose was strain-dependent. Protoplasts could regenerate on the modified R5 medium when the sucrose concentration was the same as that of culture medium used. Thus established regeneration medium provided us with regeneration ratios of 0.4 to 30%.
