细胞色素P450诱导剂对Gibberella intermedia CA3-1双羟化去氢表雄酮的影响  

Effect of cytochrome P450 inducer on the dihydroxylation of dehydroepiandrosterone by Gibberella intermedia CA3-1

作　　者：石维利 李会 史劲松 许正宏[2,3] SHI Weili;LI Hui;SHI Jinsong;XU Zhenghong(School of Pharmaceutical Science,Jiangnan University,Wuxi,214122,China;Key Laboratory of Industrial Biotechnology of the Ministry of Education,School of Biotechnology,Jiangnan University,Wuxi 214122,China;National Engineering Laboratory for Cereal Fermentation Technology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学药学院,江苏无锡214122 [2]江南大学生物工程学院工业生物技术教育部重点实验室,江苏无锡2141222 [3]江南大学粮食发酵工艺与技术国家工程实验室,江苏无锡214122

出　　处：《生物加工过程》2022年第1期29-33,共5页Chinese Journal of Bioprocess Engineering

基　　金：国家重点研发计划(2019YFA0905300);天津市合成生物学技术创新能力提升行动(TSBICIP-KJGG-001-14);江苏省高校“青蓝工程”;中央高校基本科研业务费专项资金(JUSRP221025)。

摘　　要：3β,7α,15α-三羟基雄甾-5-烯-17-酮(7α,15α-diOH-DHEA)是女用口服避孕药"优思明"主要成分屈螺酮的关键中间体。为了提高目的产物7α,15α-diOH-DHEA的摩尔得率,研究不同细胞色素P450酶(CYP450)诱导剂对Gibberella intermedia CA3-1转化去氢表雄酮(DHEA)生成7α,15α-diOH-DHEA生物转化过程的影响。结果发现:己烷、DHEA和苯可以显著提高7α,15α-diOH-DHEA的摩尔得率,其中苯是最佳诱导剂。在以上工作基础上,建立了如下的诱导过程:将进入对数生长期(接种后24 h)的种子液以10%的接种量接种到新鲜的转化培养基中,转化12 h后添加体积分数0.8%的苯进行诱导,然后在对数生长期结束时(转化24 h)将8 g/L的底物加入转化培养基进行转化。采用以上诱导条件,在5 L发酵罐中进行了生物转化。与不添加苯的工艺相比,CYP450的浓度提高了43%,7α,15α-diOH-DHEA的摩尔产率提高到68.7%±1.37%,同时转化周期缩短了8 h,这为7α,15α-diOH-DHEA的工业化生产奠定基础。3β,7α,15α-Trihydroxyandrost-5-en-17-one(7α,15α-diOH-DHEA) is a key intermediate in the synthesis of drospirenone, the active ingredient of female oral contraceptive "Yasmin". In order to increase the molar yield of the target product 7α,15α-diOH-DHEA,we studied the effect of cytochrome P450(CYP450) inducers on the biotransformation of dehydroepiandrosterone(DHEA) to 7α,15α-diOH-DHEA by Gibberella intermedia CA3-1 was investigated. Results show that as the inducers of CYP450,hexane, DHEA and benzene could significantly increase the molar yield of 7α,15α-diOH-DHEA. Among them, benzene was the best inducer. On the basis of the above work, an induction process was established as follows: the seed culture that entered the rapid growth period(inoculation for 24 h) was inoculated into a fresh transformation medium at 10% inoculation amount. Benzene 0.8%(V/V) was added for induction after 12 h of transformation, and then 8 g/L of DHEA was added to the transformation medium at the end of the logarithmic period(transformation for 24 h). With the above induction conditions, biotransformation was carried out in a 5-L fermenter. Compared with the process without the addition of benzene, the content of CYP450 was increased by 43%,the molar yield of 7α,15α-diOH-DHEA reached 68.7%±1.37%,and the conversion period was shortened by 8 h. Our findings provede for a basis for the industrial production of 7α,15α-diOH-DHEA.

关 键 词：去氢表雄酮 3β 7α 15α-三羟基雄甾-5-烯-17-酮 Gibberella intermedia CA3-1 诱导剂 苯 

分 类 号：TQ920.6[轻工技术与工程—发酵工程]