积累5-甲基四氢叶酸双抗突变株选育及条件优化.pdfVIP

82 食品与药品 Food and Drug 2010年第 12卷第03期 积累5-甲基四氢叶酸双抗突变株的选育及条件优化 1 2 1* 1 1 1 阚 静 ，胡晓川 ，卞筱泓 ，许激扬 ， 江春艳 ，沈继朵 （1. 中国药科大学生化教研室，江苏 南京210009；2. 青岛市中心医院乳腺外科，山东 青岛266042 ） 摘 要：目的 选育积累5- 甲基四氢叶酸 （5-MTHF ）的双重药物抗性高产突变株，探讨发酵时间和温度对叶酸积累形式 的影响。方法 紫外线和EMS联合诱变；磺胺、甲氨蝶呤药物培养基筛选菌株；单因素实验检测发酵条件对叶酸积累形 式的影响；HPLC检测叶酸含量和存在形式。结果 获得稳定性 良好的抗性突变株，其叶酸产量是原始菌株的2.2倍。适合 5-MTHF积累的发酵时间为16 h ，积累量为68.94 µg/g干菌。结论 抗性突变株能提高菌株产叶酸能力，5-MTHF含量受发酵 时间影响大，而发酵温度对产物形式和产量影响不显著。 关键词：抗性突变株；5- 甲基四氢叶酸；四氢叶酸；叶酸 中图分类号：Q563+.8 文献标识码：A 文章编号：1672-979X(2010)03-0082-04 Breeding of 5-Methyltetrahydrofolate-producing Double Resistance Mutant and Optimization of Accumulation Condition 1 2 1﹡ 1 1 1 KAN Jing , HU Xiao-chuan , BIAN Xiao-hong , XU Ji-yang , JIANG Chun-yan , SHEN Ji-duo (1.Dep artment of Biochemistry, China Pharmaceutical University, Nanj ing 2 10009, China; 2. Breast Surgery, Qingdao Central Hosp ital, Qingdao 266042, China) Abstract: Objective To breed 5-methyltetrahydrofolate-producing (5-MTHF-producing) double resistance mutant and investigate the contribution of fermentation time and temperature to accumulation forms of folate. Methods The original strain was induced by UV and EMS, then checked out by sulfanilamide or methotrexate medium. The influence of fermentation conditions on accumulation forms of folate was tested by single factor analysis. The content and existent form of folate were tested by HPLC. Results The double resistance mutant with stable folate production was obtained and the output of folate was 2.2-fold than that of the o