高效转化植物甾醇为9α-OH-AD的分枝杆菌诱变选育及工艺优化  

Mutation breeding of high 9α-hydroxy-androst-4-ene-3,17-dione transforming strains from phytosterols and their conversion process optimization

作　　者：马洋[1] 王向栋 王萌慧 李会[1] 史劲松[1] 许正宏[2] 

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,江苏无锡214122

出　　处：《生物工程学报》2017年第7期1198-1206,共9页Chinese Journal of Biotechnology

基　　金：国家高技术研究发展计划(863计划)(No.2011AA02A211);江苏省普通高校学术学位研究生科研创新计划项目(No.KYLX16_0826)资助~~

摘　　要：以实验室前期筛选得到的分枝杆菌(Mycobacterium sp.LY-1)为出发菌株,采用等离子诱变(ARTP)技术选育出能高效转化植物甾醇为重要甾体药物中间体9α-羟基雄甾-4-烯-3,17-二酮(9α-OH-AD)的菌株,并对其转化工艺进行优化。经过ARTP诱变选育得到遗传稳定性较好的分枝杆菌突变株Mycobacterium sp.C33,当底物植物甾醇投料浓度为15 g/L时,转化生成9α-OH-AD的摩尔得率达到15.5%,较原始菌株提高34.8%。继而采用正交实验设计方法,对突变菌的发酵培养基组分进行优化,并建立了油水双相转化体系,进一步提高了突变菌株C33的产物摩尔得率,最高达到47.0%,较优化前(15.5%)提高了2倍。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%).

关 键 词：分枝杆菌 植物甾醇 9α-OH-AD 等离子诱变 生物转化 

分 类 号：Q93[生物学—微生物学] TQ467.8[化学工程—制药化工]