Zakharova G M, Belova T S, Kamzolkina O V, Dmitrieva S V, Danilenko V N
Antibiot Khimioter. 1988 Mar;33(3):217-23.
The paper is concerned with development of conditions for cultivation of the erythromycin-producing organism and preparation and maintenance of its stable viable protoplasts. Optimal conditions for the culture growth and protoplasting were developed. Two-stage cultivation of the organism on media PB and S provided dense diffuse or diffuse local growth characterized by low differentiation and higher homogenicity. The incubation time at stage I was 66 hours and that at stage II was not more than 24 hours at respective temperatures. The culture was incubated on a shaker in the presence of glycine at the minimum concentration. The presence of glycine in the medium altered the culture cell walls which was evident from changing of staining by Gram from + to +/- . Treatment of such a culture with lysozyme for 30 minutes provided formation of up to 2.10(9) protoplasts per 1. ml. It is possible to maintain the protoplasts in frozen state at -20 degrees C in medium P for 1 month. Under such storage conditions the titer of the viable protoplasts as compared to the initial one decreased only 2-fold after the one-month storage.
本文关注产红霉素生物体的培养条件以及其稳定活原生质体的制备与保存。开发了培养生长和原生质体形成的最佳条件。在PB和S培养基上对该生物体进行两阶段培养，可实现密集的弥散状或弥散局部生长，其特点是分化程度低且同质性较高。在各自温度下，第一阶段的培养时间为66小时，第二阶段不超过24小时。培养物在摇床上于最低浓度甘氨酸存在的条件下孵育。培养基中甘氨酸的存在改变了培养物的细胞壁，这从革兰氏染色从阳性变为弱阳性可以明显看出。用溶菌酶处理这种培养物30分钟，每1毫升可形成多达2×10⁹个原生质体。可以在-20℃下将原生质体保存在培养基P中冷冻1个月。在这种储存条件下，与初始状态相比，一个月储存后活原生质体的滴度仅下降了2倍。
