Institute of Biochemistry & Physiology of Microorganisms, Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences, Federal Research Center, Pushchino, Russia.
Pharmins LTD, Pushchino, Russia.
Methods Mol Biol. 2023;2704:291-312. doi: 10.1007/978-1-0716-3385-4_18.
Engineered mutants of Mycolicibacterium spp. are known producers of valuable steroid synthons with C or C skeleton. Here we describe a method for site-directed mutagenesis of Mycolicibacterium neoaurum strains, bioconversion from phytosterol, and selective purification of C steroid 24-norchol-4-ene-3,22-dione (24-NCED) and C steroid 20-hydroxymethylpregn-4-ene-3-one (20-HMP). The yields of crystalline products with 95% purity by the method here described are 2.74 ± 0.085 g for 24-NCED and 1.42 ± 0.085 g for 20-HMP from 10 g/L phytosterol. 20-HMP is recognized as the key precursor in chemical syntheses of pharmaceutical corticosteroids and 24-NCED is a promising synthon for the synthesis of valuable steroids and own potent biological activity.
已知分枝杆菌属的工程突变体是具有 C 或 C 骨架的有价值甾体合成子的生产商。在这里，我们描述了一种分枝诺卡氏菌菌株的定点突变、从植物甾醇的生物转化以及 C 甾体 24-降胆甾-4-烯-3,22-二酮（24-NCED）和 C 甾体 20-羟甲基孕甾-4-烯-3-酮（20-HMP）的选择性纯化方法。通过本文所述方法，从 10g/L 植物甾醇中可获得纯度为 95%的结晶产物，其中 24-NCED 的产率为 2.74±0.085g，20-HMP 的产率为 1.42±0.085g。20-HMP 被认为是化学合成药物皮质甾类的关键前体，而 24-NCED 是合成有价值的甾体和具有自身强大生物活性的合成子的有前途的前体。
