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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2017, Vol. 37 Issue (3): 73-77    DOI: 10.13523/j.cb.20170310
技术与方法     
Gibberella intermedia C2转化4-雄甾烯-3、17-二酮的研究
姚韧辉1, 董卓1, 李会2
1. 唐山职业技术学院 唐山 063000;
2. 江南大学药学院 无锡 214122
Biotransformation of Androst-4-en-3,17-dione by Gibberella intermedia C2
YAO Ren-hui1, DONG Zhuo1, LI Hui2
1. Tangshan Vocational & Technical College, Tangshan 063000, China;
2. School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, China
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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-OH-AD,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%。

关键词: 赤霉菌生物转化结构鉴定4-雄甾烯-3、17-二酮工艺优化    
Abstract:

Steroids with unique biological activity are widely used as anti-inflammatory, diuretic, anti-androgenic, 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-diOH-AD 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-diOH-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 250ml shake flask with 30ml medium at 28℃, substrate concentration 6.0g/L, culture for 24h and biotransformation for 84h at 220r/min. The molar conversion reached 81.5% under above conditions.

Key words: Biotransformation    Process optimization    Androst-4-en-3, 17-dione    Gibberella intermedia    Structure identification
收稿日期: 2016-10-11 出版日期: 2017-03-25
ZTFLH:  TQ920  
通讯作者: 李会     E-mail: huilijn@163.com
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引用本文:

姚韧辉, 董卓, 李会. Gibberella intermedia C2转化4-雄甾烯-3、17-二酮的研究[J]. 中国生物工程杂志, 2017, 37(3): 73-77.

YAO Ren-hui, DONG Zhuo, LI Hui. Biotransformation of Androst-4-en-3,17-dione by Gibberella intermedia C2. China Biotechnology, 2017, 37(3): 73-77.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170310        https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I3/73

[1] Mahato S B, Garai S. Advances in microbial steroid biotransformation. Steroids, 1997, 62(4):332-345.
[2] Baker M E. Origin and diversification of steroids:co-evolution of enzymes and nuclear receptors. Molecular and Cellular Endocrinology, 2011, 334(1-2):14-20.
[3] Weber A, Kennekke M, Klages U, et al. Process for the Production of 17-oxosteroids Via the Ermentative Oxidation of 17β-hydroxysteroids by Mycobacterium:US, US6203493A. 1993-05-17.
[4] Roberts S M, Eddolls J P, Willetts A J, et al. Preparation of Adrenocorticoid Steroids:United Kingdom, GB2318790. 1997-10-31.
[5] Tong W Y, Dong X. Microbial biotransformation:recent developments on steroid drugs. Recent Patents on Biotechnology, 2009, 3(2):141-153.
[6] Peart P C, McCook K P, Russell F A, et al. Hydroxylation of steroids by Fusarium oxysporum, Xophiala jeanselmei and Ceratocystis paradoxa. Steroids, 2011, 76(12):1317-1330.
[7] Li H, Liu H M, Ge W, et al. Synthesis of 7α-hydroxy-dehydroepiandrosterone and β-hydroxy-dehydroepiandrosterone. Steroids, 2005, 70(14):970-973.
[8] Lamm A S, Chen A R M, Reynolds W F, et al. Steroid hydroxylation by Whetzelinia sclerotiorum, Hanerochaete chrysosporium and Mucor plumbeus. Steroids, 2007, 72(9-10):713-722.
[9] Choudhary M I, Erum S, Atif M, et al. Biotransformation of 20S)-20-hydroxymethylpregna-1,4-dien-3-one by four filamentous fung. Steroids, 2011, 76(12):1288-1296.
[10] Donova M V, Egorova O V. Microbial steroid transformations:current state and prospects. Applied Microbiology and Biotechnology, 2012, 94(6):1423-1447.
[11] Choudhary M I, Mohammoad M Y, Musharraf S G, et al. New oxandrolone derivatives by biotransformation using Rhizopus stolonife. Steroids, 2009, 74(13-14):1040-1044.
[12] Koshimura M, Utsukihara T, Hara A, et al. Hydroxylation of steroid compounds by Gelasinospora retispora. Journal of Molecular Catalysis B:Enzymatic, 2010, 67(1-2):72-77.
[13] Kollerov V V, Shutov A A, Fokina V V, et al. Bioconversion of C19- and C21-steroids with parent and mutant strains of Curvularia lunata. Applied Biochemistry and Microbiology, 2010, 46(2):198-205.
[14] Huang L H, Li J, Xu G, et al. Biotransformation of dehydroepiandrosterone (DHEA) with Penicillium griseopurpureum Smith and Penicillium glabrum (Wehmer) Westling. Steroids, 2010, 75(13-14):1039-1046.
[15] Wu Y, Li H, Zhang X M, et al. Efficient hydroxylation of functionalized steroids by Colletotrichum lini ST-1. Journal of Molecular Catalysis B:Enzymatic, 2015, 120:111-118.

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