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
投稿时间:2016-01-13  
DOI:  10.13345/j.cjb.170014
中文关键词:分枝杆菌,植物甾醇,9α-OH-AD,等离子诱变,生物转化
英文关键词:Mycobacterium sp., phytosterol, 9α-OH-AD, atmospheric and room temperature plasma, biotransformation
基金项目: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。
作者单位E-mail
马洋 1 江南大学 药学院江苏 无锡 214122  
王向栋 1 江南大学 药学院江苏 无锡 214122  
王萌慧 1 江南大学 药学院江苏 无锡 214122  
李会 1 江南大学 药学院江苏 无锡 214122 lihui@jiangnan.edu.cn 
史劲松 1 江南大学 药学院江苏 无锡 214122  
许正宏 2 江南大学 生物工程学院江苏 无锡 214122  
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中文摘要:
      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。
英文摘要:
      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%).
马洋,王向栋,王萌慧,李会,史劲松,许正宏.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[J].生物工程学报,2017,33(7):1198~1206
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