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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%。
Spiropreussione A (SP-A) is a metabolite of the endophytic fungus Preussia sp. From AS-5. In vitro experiments showed that the half-inhibitory concentration (IC50) of SP-A to human ovarian cancer cell A2780 and human hepatocellular carcinoma cell BEL-7404 were 2.4 and 3.0μmol / L, respectively. Based on the content of SP-A and the biomass of AS-5, single factor experiment and orthogonal experiment were used to optimize the AS-5 liquid fermentation medium and culture conditions for SP-A accumulation. The results showed that the optimal culture medium for the production of SP-A by AS-5 fermentation was 2% glucose, 3% wheat bran, 0.3% potassium dihydrogen phosphate, 0.15% magnesium sulfate and pH 7.0. The conditions were 250mL shake flask 125mL medium, inoculated 6 diameter 9mm PDA strain, culture temperature 25 ℃, fermentation period 16d. Under this condition, the content of SP-A reached (25.02 ± 1.02) mg / bottle, which was 46.5% higher than that before optimization [(17.08 ± 3.24) mg / bottle].