生物法制备三羟基雄甾烯酮的分离提取工艺研究  

Separation and Extraction Process of 3β,7α,15α-trihydroxy-5-androsten-17-one in Biological Preparation

作　　者：倪瑜 尹思淇 李会 史劲松 许正宏[2] NI Yu;YIN Si-qi;LI Hui;SHI Jin-song;XU Zheng-hong(School of Medicine,Jiangnan University,Wuxi 214122;School ofBiotechnology,Jiangnan University,Wuxi 214122)

机构地区：[1]江南大学药学院,无锡214122 [2]江南大学生物工程学院,无锡214122

出　　处：《生物技术通报》2018年第8期67-74,共8页Biotechnology Bulletin

基　　金：国家"863计划"重大项目(2011AA02A211)

摘　　要：三羟基雄甾烯酮(7α,15α-diOH-DHEA)是第四代女用口服避孕药"优思明"有效成分屈螺酮的关键中间体,具有重要的市场价值。采用生物转化法可以将底物去氢表雄酮(DHEA)转化成7α,15α-diOH-DHEA,该方法具有转化效率高、环境友好等优势。本研究对发酵液中产物的分离提取工艺进行了优化,确定了三羟基雄甾烯酮最适的分离提取工艺。结果表明,添加4%(V/V)的氢氧化钠-乙醇溶液预处理,发酵液上清和沉淀分别用1.5倍和2倍的乙酸乙酯和乙醇萃取,洗脱剂选择二氯甲烷:乙醇=15∶1,产物分离后纯度达98.1%,纯化收率可达91.4%。通过质谱(MS),红外光谱(IR)以及氢谱(1H-NMR)对产物结构进行鉴定,与7α,15α-diOH-DHEA标准品结构一致。3β,7α, 15α-trihydroxy-5-androsten-17-one （7α, 15α-diOH-DHEA ） is a key intermediate of drospirenone, the active ingredient of the fourth generation of oral contraceptive Yasmin for women, thus it has the significant value in the market. The substrate dehydroepiandrosterone （ DHEA ） can be converted to 7α, 15α-diOH-DHEA through biotransformation, which has advantages of high conversion rate and environmentally friendly. In this study, the extraction and separation of the product in the fermentation broth were optimized, and the optimal process of extracting and separating 7α, 15α-diOH-DHEA were determined. Results showed that fermentation broth was pretreated with 4% （ V/V ） sodium hydroxide-ethanol, and its supernatant and sediment were extracted with 1.5 times and 2 times of ethyl acetate and ethanol respectively. When dichloromethane ： ethanol = 15 ： 1 was select as eluent, the purity of product reached 98.1% after separation, and the yield of purification reached 91.4%. The structure of the product was characterized by MS, IR and 1H-NMR, and it was consistent with the structure of standard 7α, 15α-diOH-DHEA.

关 键 词：三羟基雄甾烯酮 亚麻刺盘孢ST-1 去氢表雄酮 提取 分离纯化 

分 类 号：TQ460.6[化学工程—制药化工] TQ920.6[轻工技术与工程—发酵工程]