Blakeslea trispora H_1^-原生质体制备与再生条件研究

Study on formation and regeneration of protoplast from Blakeslea trispora H_1^-

以β-胡萝卜素生产菌株Blakeslea trisporaH1-为研究对象,对其原生质体形成及再生条件进行了研究。得知最佳形成与再生条件为:以摇瓶方式培养菌丝体,培养菌龄为56h,酶解温度为30℃,酶质量分数2.0%,混合酶组成为m(蜗牛酶)∶m(纤维素酶)=6∶4,酶解时间为120min,渗稳剂为0.6mol/L MgSO4·7H2O。Blakeslea trisporaH1-原生质体形成数为7.9×106个/mL,再生率为3.9%。

The formation and regeneration of the protoplast from Blakeslea trispora H1^- were studied, which is a β-carotene producing strain. The results showed that the optimal conditions are as follows： shaking culture as the mycelium culture method, the mycelium cultured 56 h as operated material, 30 ℃ as hydrolysis temperature, 120 min as hydrolysis time, 2.0% mixed enzyme re（snail enzyme） ： m（cellulose enzyme）=6 ： 4 enzyme system, 0.6 mol/L MgSO4 · 7H2O as osmotic stabilizer. Under these conditions, the formation rate of the protoplast is up to 7.9×10^6/mL, and the generation rate is up to 3.9%.