Biotechnology Bulletin ›› 2018, Vol. 34 ›› Issue (8): 67-74.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0106

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

NI Yu1, YIN Si-qi1, LI Hui1, SHI Jin-song1, XU Zheng-hong2   

  1. 1. School of Medicine,Jiangnan University,Wuxi 214122;
    2. School of Biotechnology,Jiangnan University,Wuxi 214122
  • Received:2018-01-30 Online:2018-08-26 Published:2018-09-04

Abstract:

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 and1H-NMR,and it was consistent with the structure of standard 7α,15α-diOH-DHEA.

Key words: 7a,15a-diOH-DHEA, Colletotrichumlini ST-1, DHEA, extraction, separation and purification