Gibberella intermedia C2转化4-雄甾烯-3、17-二酮的研究  

Biotransformation of Androst-4-en-3,17-dione by Gibberella intermedia C2

作　　者：姚韧辉[1] 董卓[1] 李会 

机构地区：[1]唐山职业技术学院,唐山063000 [2]江南大学药学院,无锡214122

出　　处：《中国生物工程杂志》2017年第3期73-77,共5页China Biotechnology

摘　　要：甾体化合物具有独特的生理活性,已被广泛应用于抗炎、利尿、免疫、避孕及抗癌等领域。近些年,生物催化与转化在甾体药物中间体合成中发挥的作用日益强大。为了能够合成一些具有潜在价值的新型甾体化合物,以实验室菌种库中保藏的一株Gibberella intermedia C2为研究对象,选取了雄甾烷中一种有广泛用途的化合物4-雄甾烯-3、17-二酮(简称雄烯二酮,AD)为底物进行生物转化。转化液经提取分离,最终获得2个转化产物,经结构鉴定分别为15α-OH-AD和11α,15α-diOH-AD。转化机制研究发现,G.intermedia C2先将底物的15位羟基化生成15α-OHAD,再将其11位羟基化形成双羟基产物。赤霉菌能够特异性、有序地完成对AD的两步羟化反应。此外,通过工艺优化,确定了羟化4AD反应的最适工艺参数如下:发酵培养基的初始pH 6.5,装液量30ml/250ml,底物浓度6.0g/L,转化温度28℃,摇床转速220r/min,转化周期为84h。此时,底物AD的摩尔转化率达到81.5%。Steroids with unique biological activity are widely used as anti-inflammatory,diuretic,antiandrogenic,contraceptive and anti-cancer agents. In recent years,the role of biocatalysis and biotransformation in the synthesis of steroid drug intermediates is becoming more and more significant. In order to synthesize the potential steroidal compounds, biotransformation of androst-4-en-3,17-dione（ androstenedione, AD） by Gibberella intermedia C2 was investigated. The two products was identified as 15α-OH-AD and 11α,15α-diOHAD by structure identification. The results of the conversion mechanism research showed that androstenedione is first converted into 15α-OH-AD with the 15α-hydroxylation of AD,and subsequently,into 11α,15α-di OH-AD after the 11α-hydroxylation of 15α-OH-AD. G. intermedia C2 can selectively and sequentially perform two DHEA hydroxylation reactions. In addition,the optimum biotransformation conditions of androstenedione were obtained as follows： initial pH value 6. 5,the strain incubated in 250 ml shake flask with 30 ml medium at 28℃,substrate concentration 6. 0g/L,culture for 24 h and biotransformation for 84 h at 220r/min. The molar conversion reached 81. 5% under above conditions.

关 键 词：4-雄甾烯-3、17-二酮 赤霉菌 生物转化 结构鉴定 工艺优化 

分 类 号：TQ920[轻工技术与工程—发酵工程]