Formation of Streptomyces protoplasts during cultivation in liquid media with lytic enzyme

Many streptomycetes strains are hardly or not at all transformable via protoplasts, there is a problem with the regeneration of protoplasts. We found that protoplasts formed directly in cultivation media under submerged conditions presence lytic enzyme. Actinophage ?1/6 endolysin and lysozyme were used this study. Streptomyces cultivated several glycine enzyme for 24 48h. The highest amounts (about 3 x 107 cfu/ml medium) together (95%) transformation frequency 2 106 – cfu/?g DNA) obtained reproducibly YEME medium high sucrose content. S. aureofaciens B96, as strain because difficulties protoplast preparation their further regeneration, was same procedure applied to lividans 66 TK24 coelicolor A3(2), model strains, confirm general use method. Moreover, such process appropriate additional quick isolation either chromosomal well plasmid DNA could be recombinant techniques.