内生真菌gyzy-20生产高三尖杉酯碱的成分分析及条件优化

Components Analysis of Homoharringtonine Produced by Endophytic Fungi Strain gyzy-20 and Optimization for Fermentation Condition

[目的]为充分发掘篦子三尖杉(Cephalotaxus oliveriMast)内生真菌的应用价值。[方法]采用HPLC对内生真菌菌株gyzy-20的发酵提取组分进行鉴定,并采用正交试验进一步优化生产工艺。[结果]菌株gyzy-20的代谢产物中有高三尖杉酯碱生成,提取时间对高三尖杉酯碱含量的影响较大。优化的生产工艺为:碳源为葡萄糖,发酵10 d后超声5 min,添加2倍体积二氯甲烷萃取3 h,优化后的发酵和提取方法可提高高三尖杉酯碱的含量1.5倍以上,达到0.26μg/ml。[结论]该研究可为利用微生物发酵生产HHT提供可能。

[Objective]In order to explore the application value of endophytic fungi isolated from Cephalotaxus oliveri Mast.[Method]The fermentation products of strain gyzy-20 were extracted and analyzed by HPLC,and then the producing process was optimized by orthogonal experiment.[Result]The metabolites of strain gyzy-20 contained homoharringtonine（HHT）,in which the HHT content was affected by extraction time largely.The optimized producing process was as follow：C sources glucose,ultrasonic extraction 5 min after fermenting 10 d,extraction 3 h after adding 2 times volumes of dichloromethane.The HHT content up to 0.26 μg/ml of fermentation broth,increased more than 1.5 times through optimized producing process comparing with that of before.[Conclusion]The study can provide probability on HHT production through microbe fermentation.