[Preparation and characteristics of protoplasts of Streptomyces kanamyceticus]. / Poluchenie i kharakteristika protoplastov shtamma Streptomyces kanamyceticus.

To prepare actively regenerating protoplasts of S. kanamyceticus, the influence of the conditions of the mycelium cultivation, the culture age, lytic conditions, composition of the regeneration medium, the procedure of the culture inoculation to the regeneration medium and other parameters were studied. The study resulted in development of optimal conditions for preparation of S. kanamyceticus protoplasts in a number of 1.10(9) protoplasts per ml. The cultivation on the ST medium with 10 to 15% sucrose and addition of glycine up to 1% for 30 hours (the stationary growth phase) followed by treatment of the culture with lysozyme in an amount of 2 mg/ml for 1 hour at 32 degrees C provided preparation of up to 100% of actively regenerating protoplasts free of mycelium fragments. The size of the protoplasts increased up to 1.5 micron against the usually observed size of 0.7 to 1.0 micron with using modified lyzing buffer with 20% of sucrose according to the method recommended for S. erythreus. However, 50 to 70% of the protoplasts had point of linear regions in the cell walls, which suggested that spheroplasts were mainly forming and the phenomenon was associated with the characteristic properties of the strain cell wall structure.

[Development of protoplasts of Streptomycetes on media favoring the regeneration and formation of L-forms]. / Razvitie protoplastov streptomitsetov na sredakh, sposobstvuiushchikh regeneratsii i obrazovaniiu L-form.

The protoplasts of three Streptomyces species and their regenerative ability were studied using light microscopy. When Streptomyces lividans and S. erythraeus protoplasts are cultivated on regeneration media, their regeneration is not synchronous during the first day; some protoplasts revert to yield the mycelial form and also L-forms of these cultures are produced. If the protoplasts are transferred to a medium inducing L-forms, they grow and multiply for a long time with the production of L-form colonies. This process is maintained if S. lividans L-form cells are passaged on the medium inducing L-forms, but the protoplasts revert to yield the mycelial form on the regeneration medium.