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

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

作　　者：陈晓梅[1] 施琦渊[1] 王春兰[1] 李兵[1] 王爱荣[1] 郭顺星[1] 

机构地区：[1]中国医学科学院北京协和医科大学药用植物研究所,北京100193

出　　处：《菌物学报》2013年第4期729-740,共12页Mycosystema

基　　金：国家自然科学基金项目“一种真菌抗艾滋病病毒(HIV)活性成分研究”(No.31170016);科技部“重大新药创制”专项“创新药物候选物研究”(No.2012ZX09301-002-001)

摘　　要：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）, a metabolite of endophytic fungus Preussia sp. AS-5, showed cytotoxicity in vitro experiment toward A2780 and BEL-7404 cells with ICs0 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%, KH2PO4 0.3%, and MgSO4 0.15%; the initial pH was 7.0. The optimum fermentation conditions were： inoculating 6 pieces of PDA fungal plates （Ф9mm） in 250mL Edenmeyer 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）rag/flask].

关 键 词：内生真菌 代谢产物 抗肿瘤活性 发酵条件优化 

分 类 号：R284[医药卫生—中药学]