论文摘要
CDP-choline 是磷脂代谢的重要前体,广泛用于治疗各种脑血管疾病。本文选育了生物转化CDP-choline 的菌株产朊假丝酵母candida utilis,研究了发酵条件及发酵培养基,探索了其酶反应体系的优化条件。利用放线菌酮和TTC 作为一种筛选压力,应用紫外线及N+注入等方法对candida utilisYL 进行了诱变筛选。结果表明,突变株candida utilis YL-2-24 的CDP-choline 比出发菌株candida utilis YL 提高2.01 倍。对candida utilis YL-2-24 的发酵工艺进行了优化。通过培养基的单因子试验及正交试验,最终确定最佳发酵培养基配方为蔗糖4%、蛋白胨1.0%、NaNO32.0%、K2HPO40.5%、MgSO4·7H2O 0.5%、KH2PO4 0.1%;摇瓶发酵条件为初始pH5.5,接种量10%,装液量25ml/250ml , 温度32℃, 培养时间24h , 转速230 r/min , 生物转化生成18.63mg/gCDP-choline。candida utilis YL-2-24 细胞转化的酶促反应条件为CMP10mM、磷酸盐浓度300mM、磷酸胆碱50mM、葡糖糖1000mM、34℃、PH7.4、湿细胞的加量为600mg/ml、反应时间为10h,生物转化生成CDP-choline 的得率为28.63%,转化率达到84.4%。
论文目录
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