褐黄孢链霉菌原生质体制备与再生

Preparation and Regeneration of Protoplast from Streptomyces gilvosporeus

以纳他霉素产生菌株褐黄孢链霉菌SG-2002为出发菌株,考察了菌丝体培养基、甘氨酸浓度、培养时间、溶菌酶浓度、酶解温度、酶解时间及再生培养基对原生质体形成与再生的影响。原生质体形成和再生的最佳条件为S培养基中添加1%的甘氨酸,菌丝体培养30h,溶菌酶浓度40mg/(g菌体干重),酶解温度30℃,酶解时间60min。褐黄孢链霉菌的原生质体形成量达到4.0×106个/mL,再生培养基选择R5′培养基,再生率为9.0%。

Natamycin-producing Streptomyces gilvosporeus strain SG-2002 was taken as starting strain to observe the effects of mycelial medium, glycine concentration, culture time, lysozyme concentration, enzymolytic temperature, enzymolytic time, and regeneration medium on protoplast formation and regeneration. The optimum conditions for protoplast formation and regeneration were as follows ： S medium adding 1% of glycine, mycelium culture for 30 h, lysozyme concentration 40 mg/g （ DWC ）, lysozymc temperature 30 ℃ and lysozyme treating time 60 rain. Under these conditions, the number of protoplast reached up to 4.0 × 10^6 cfu/mL. The optimum medium of protoplast regeneration is R5 and the regeneration rate was 9.0%.