内生真菌Preussia sp.液体发酵生产抗肿瘤活性产物Spiropreussione A 的研究

螺光黑壳菌酮A(Spiropreussione A,SP-A)是编号为AS-5的光黑壳属内生真菌Preussia sp.的代谢产物。体外实验表明SP-A对人卵巢癌细胞A2780和人肝癌细胞BEL-7404的半数抑制浓度(IC50)分别为2.4和3.0μmol/L。以SP-A的含量为主要指标,结合AS-5的生物量,通过单因素实验和正交实验,优化确定了适合SP-A积累的AS-5液体发酵培养基和培养条件。研究结果表明:AS-5发酵生产SP-A的最优培养基为葡萄糖2%,麦麸3%,磷酸二氢钾0.3%,硫酸镁0.15%,pH7.0;该菌株最佳摇瓶发酵条件为250mL摇瓶装125mL培养基,接种6片直径9mm的PDA菌片,培养温度25℃,发酵周期16d。在此条件下发酵,SP-A的含量可以达到(25.02±1.02)mg/瓶,比优化前的含量(17.08±3.24)mg/瓶]提高了46.5%。

The production of Spiropreussione A, an antitumor metabolite produced by endophytic fungus Preussia sp., with liquid fermentation

Spiropreussione A (SP-A), a metabolite of endophytic fungus Preussia sp. AS-5, showed cytotoxicity in vitro experiment toward A2780 and BEL-7404 cells with IC 50 values of 2.4 and 3.0μmol/L, respectively. The objective of this research was to optimize the medium and the liquid fermentation condition of isolate AS-5 for the production of SP-A by single factor and orthogonal tests. The results showed that the best medium for SP-A production contained glucose 2%, wheat bran 3%, KH 2 PO 4 0.3%, and MgSO 4 0.15%; the initial pH was 7.0. The optimum fermentation conditions were: inoculating 6 pieces of PDA fungal plates (Φ9mm) in 250mL Erlenmeyer flask containing 125mL medium, and incubating the fungus under the temperature of 25℃ for 16 days. Under such a condition, the content of SP-A reached (25.02±1.02)mg/flask and increased by 46.5% as compared to that before optimization (17.08±3.24)mg/flask].