生物素与氨基酸对林可霉素生物合成的影响.docVIP

生物素与氨基酸对林可霉素生物合成的影响 【摘要】 在合成培养基中利用林可链霉菌发酵生产林可霉素。当向培养基中加入生物素和氨基酸时，林可霉素的产量受到很大影响。本研究中首先采用两水平因子设计法筛选出6个显著影响因子，即生物素、谷氨酸、缬氨酸、蛋氨酸、亮氨酸和酪氨酸；然后采用中心组合设计法，得到上述6因子的优化含量分别为30μg/L、100.5、83、29、117.5和58.5mg/L；最后在摇瓶中进行验证实验，优化条件下发酵液的生物效价为2116μg/ml；对照样品I(培养基中无生物素和氨基酸)和II(培养基中按原配方加入6个显著影响因子)的生物效价分别为893和1481μg/ml，与样品I和II比较，生物效价分别提高136.62%和42.88%。 【关键词】 林可链霉菌； 林可霉素； 发酵； 生物合成； 响应面法 Effects of biotin and amino acids on biosynthesis of lincomycin Li Xiao1, Chu Ju1, Zhang Siliang1, Hang Haifeng1, Zhuang Yingping1 and Ge Youqun2 (1 State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237; 2 Jiangxi Guoyao Pharmaceutical Co.,Ltd, Nanchang 330052) ABSTRACT Streptomyces lincolnensis was utilized to produce lincomycin by fermentation in chemically defined medium, enormous effect on the yield of lincomycin would occur when biotin and amino acids were added to the medium. In this research, firstly, 6 significant factors were found with two level factorial design, namely Dbiotin, Lglutamic acid, Lvaline, Lmethionine, Lleucine and Ltyrosine; secondly, the optimized ingredients of 6 factors mentional above were obtained with central composite design, they were 30μg/L and 100.5, 83, 29, 117.5 and 58.5mg/L respectively; finally, verification test was accomplished in shaking flasks, the titre of the fermented broth was 2116μg/ml on the optimized conditions, but the titres of the control I (without Dbiotin and amino acids in the medium ) and control II (adding 6 significant factors to medium according to original ingredients) were 893μg/ml and 1481μg/ml respectively, compaired with control I or II, increased by 136.62% and 42.88%. KEY WORDS Streptomyces lincolnensis; Lincomycin; Fermentation; Biosynthesis; Response surface method 林可霉素是一种高效广谱抗生素，临床用于由抗革兰阳性菌引起疾病的治疗［1］。林可霉素A的分子式为C18H34N2O6S，分子量为406.56，它与林可霉素B在结构上的区别［2，3］为：林可霉素A 4位上为正丙基，而林可霉素B4位上是乙基， 两者在药理上存在着很大的区别，林可霉素B的抑菌活性比林可霉素A低，但对人的毒副