摘要
[目的]探讨米曲霉原生质体制备的高效方法。[方法]统计米曲霉菌丝体检测不同生长时间(72、84、96、108、120 h)的菌丝、不同渗透压(分别使用0.8 mol/L蔗糖、氯化钠和葡萄糖溶液作为渗透压稳定剂)和不同组合酶系(溶菌酶、纤维素酶和蜗牛酶、混合酶系的配比按照L9(34)正交设计)下的原生质体产量。[结果]经检测,培养108 h菌丝体、0.8 mol/LNaCl渗透压稳定剂原生质体产量较高。同时溶菌酶(2 mg/ml) +纤维素酶(6 mg/ml) +蜗牛酶(6 mg/ml)组合酶系原生质体产量较高,达9×105个/ml。[结论]原生质体的产量受菌体自身状况和外界条件影响。该研究为大量制备高活力的原生质体及进行细胞融合试验奠定了基础。
[Objective] To explore the efficient way for the preparation of Aspergillus oryzae protoplast.[Method] The statistics of protoplast yield by the enzymolysis of mycelia with different culture time,different osmotic stabilizers and different concentrations of enzyme system was carried out in this study.[Result] The protoplast yield was higher when the mycelium was cultivated for 108 h,and the osmotic stabilizer was 0.8 mol/L of NaCl.With the enzymes system of 2 mg/ml of lysozyme,6 mg/ml of cellulose and 6 mg/ml of glusulase,the protoplast yield achieved its highest,which was up to 9.0×105 per ml.[Conclusion] The yield of protoplast was affected by the situation of mycelia themselves and external conditions.This study had provided a basis for the preparation of a large number of active protoplasts and the further researches on cell fusion.
关键词
米曲霉
原生质体
酶解
Aspergiilus oryzae
Protoplast
Enzymolysis
