Key Laboratory of Industrial Fermentation Microbiology (Tianjin University of Science and Technology), Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, Tianjin Engineering Research Center of Microbial Metabolism and Fermentation Process Control, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, People's Republic of China.
Bioprocess Biosyst Eng. 2019 Jun;42(6):933-940. doi: 10.1007/s00449-019-02092-y. Epub 2019 Mar 8.
Boldenone (BD) is an important steroid hormone drug which is the derivative of testosterone. In this study, an ordered biotransformation method was proposed employing Arthrobacter simplex and recombinant Pichia pastoris with 17β-hydroxysteroid dehydrogenase from Saccharomyces cerevisiae to produce BD from androst-4-ene-3,17-dione (AD) efficiently. To lower the oxidation towards BD in A. simplex, the transformation was conducted sequentially by C1,2 dehydrogenation in A. simplex and 17β-carbonyl reduction in recombinant P. pastoris GS115. Moreover, the reaction system was inactivated before recombinant P. pastoris GS115 was added to further inhibit the oxidation of BD by A. simplex, and the productivity of BD was improved 10.6% compared with the control. Furthermore, by optimizing the conditions of transformation from AD to BD, 4.2 g/L BD was generated with 83% productivity from 5.0 g/L AD, which was the highest productivity reported by biological method. This study offers a promising method to produce BD by ordered biotransformation system, which can also be used to manufacture other steroidal compounds that are difficult to acquire directly.
宝丹酮（BD）是一种重要的甾体激素药物，是睾酮的衍生物。本研究采用阿氏节杆菌和重组毕赤酵母，利用酿酒酵母中的 17β-羟甾脱氢酶，提出了一种有序生物转化方法，可从雄甾-4-烯-3,17-二酮（AD）高效生产 BD。为降低阿氏节杆菌中 BD 的氧化，通过 C1,2 脱氢在阿氏节杆菌中，17β-羰基还原在重组毕赤酵母 GS115 中依次进行转化。此外，在添加重组毕赤酵母 GS115 之前，使反应体系失活，进一步抑制阿氏节杆菌对 BD 的氧化，与对照相比，BD 的产率提高了 10.6%。此外，通过优化从 AD 到 BD 的转化条件，从 5.0 g/L AD 生成了 4.2 g/L 的 BD，产率为 83%，这是生物法报道的最高产率。本研究提供了一种通过有序生物转化系统生产 BD 的有前途的方法，该方法也可用于制造其他难以直接获得的甾体化合物。
