[以植物甾醇为原料的9α-羟基-雄甾-4-烯-3,17-二酮高产转化菌株的诱变育种及其转化工艺优化]
[Mutation breeding of high 9α-hydroxy-androst-4-ene-3,17- dione transforming strains from phytosterols and their conversion process optimization].
作者信息
Ma Yang, Wang Xiangdong, Wang Menghui, Li Hui, Shi Jinsong, Xu Zhenghong
机构信息
School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, Jiangsu, China.
School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
出版信息
Sheng Wu Gong Cheng Xue Bao. 2017 Jul 25;33(7):1198-1206. doi: 10.13345/j.cjb.170014.
In order to improve transformation efficiency of phytosterols into 9α-hydroxylation of 4-androstene-3,17-dione (9α-OH-AD) by Mycobacterium sp. LY-1, we studied the strains breeding using atmospheric and room temperature plasma (ARTP) technology and optimized their conversion process. A high production strain named C33 with a good genetic stability was selected and the product molar yield reached to 15.5%, 34.8% higher than that of original strain with 15 g/L phytosterols. Furthermore, the fermentation medium was optimized through the design of orthogonal experiment. Besides, oil-water bidirectional transformation system was set up to improve the 9α-OH-AD molar yield of mutant strain C33. With adding 12 mL soybean oil to each 1 g phytosterols, the molar yield of 9α-OH-AD reached 47.0%, which increased twice than that of control (15.5%).
为提高分枝杆菌属LY-1菌株将植物甾醇转化为4-雄烯二酮-3,17-二酮9α-羟基化产物（9α-OH-AD）的效率，我们研究了利用常压室温等离子体（ARTP）技术进行菌株选育，并优化其转化过程。筛选出一株遗传稳定性良好的高产菌株C33，在15 g/L植物甾醇条件下，产物摩尔产率达到15.5%，比原始菌株提高了34.8%。此外，通过正交试验设计优化了发酵培养基。另外，建立了油水双向转化体系以提高突变菌株C33的9α-OH-AD摩尔产率。每1 g植物甾醇添加12 mL大豆油时，9α-OH-AD的摩尔产率达到47.0%，比对照（15.5%）提高了两倍。
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