泰妙菌素生产菌种原生质体制备条件优化

Optimization on Protoplast Preparation Conditions of Tiamulin Producing Strain

[目的]探讨泰妙菌素生产菌种原生质体的最优制备方法。[方法]通过对原生质体制备条件进行优化,建立泰妙生产菌种原生质体制备体系。[结果]优化后制备条件：以培养3 d的菌丝体为材料,将其置于含质量浓度0.10%溶壁酶、0.60 mol/L MgSO_4的酶液中,25℃酶解3.0~3.5 h,将酶解液用G2砂芯漏斗过滤,滤液即为原生质体悬液。采用该方法,泰妙生产菌种原生质体制备率达1.5×10~6个/m L。[结论]试验结果为提高泰妙菌素生产菌种的产量提供了理论依据。

[Objective]The aim was to explore the optimal protoplast preparation method of Tiamulin producing strain.[Method]We established a system for protoplast preparation of Tiamulin producing strain by optimizing protoplast preparation conditions. [Result]The opotimal conditions were as followed： used 3 days cultured mycelium as material; put it into a solution which contain 0. 10% lyticase,0. 60 mol/L MgSO_4; hydrolyzed the mycelium with enzyme under 25 ℃ for 3. 0-3. 5 hours,then filtered the solution with G2 sintered discs; the filter liquor was protoplast suspension. By using this method,the protoplast preparation rate of Tiamulin producing strain could reach 1. 5 × 10~6 per milliliter.[Conclusion]The results provide theoretical basis for the improvement of Tiamulin producing strain.