比较2种不同优化方法对分离纯化梨孢镰孢菌(Fusarium poae)产生的T-2毒素的效果,并得到高纯度的T.2毒素,解决国内T-2毒素产业化问题。在优化Fusariumpoae产毒培养条件的基础上,对Burmeister的提取方法和Gregory培养基进一步优化以最大限度地提高T-2毒素的产率,从而得到高纯度并且高产率的T-2毒素。目标菌株最适产毒培养基成分:NH4H2PO4 1g,KCl 0.2g,MgSO4·7H2O 0.2g,葡萄糖10g,酵母膏5g,CuSO4·5H2O 0.005g,ZnSO4·7H2O 0.01g,H2O 1000mL。最佳产毒培养条件为8—25℃间隔12h变温、前期光照后期黑暗、前期振荡后期静止的培养条件下培养28d。在这种条件下可提高T-2毒素的产率达5倍。用丙酮氯仿进行分离,免疫亲和拄进行纯化得到高纯的T-2毒素。优化后的Burmeister提取方法提高了T-2毒素的产率。
The effects to isolate and purify T-2 toxin produced by Fusarium poae (FP) with two different optimized methods were compared and obtained high purity of T-2 toxin to solve its industrialization problem in China. Based on the optimization of toxin producing conditions by FP, Burmeister extraction and Gregory medium were further opti- mized in order to improve the productivity of T-2 toxin to the maximum thereby to obtain high purity and high productivity of T-2 toxin. The results showed that the most suitable toxin producing medium components of target strain were as follows: NH4H2PO4 1 g, KCl 0.2 g, MgSO4 - 7H20 0.2 g, glucose 10 g, yeast extract 5 g, CuSO4 · 5H2O 0. 005 g, ZnSO4 · 7H2O 0.01 g, H2O 1 000 mL. The most optimized conditions to produce toxin was euhured for 28 days at interval 8 - 25 ℃ of poikilothermia for 12 hours under illumination and shaking at early stage, then shading and culturing at rest at later stage, the production of T-2 toxin reached to maximal level of 5 mg, which was five times more than before. After separated by acetone-chloroform and purified by immune affinity column, and obtained high purity T-2 toxin. The optimized Burmeister extraction improved the productivity of T-2 toxin