增加aveBⅣ基因拷贝数对于表柔红霉素产生菌发酵单位的影响  

Influence of Increase in the Number of aveB Ⅳ Gene Copy on Improving the Fermentation Unit of Epidaunorubicin-Producing Strain

作　　者：李继安[1] 邵雷[1] 景兰[1] 陈代杰[1] 朱宝泉[1] 

机构地区：[1]上海医药工业研究院创新药物与制药工艺国家重点实验室(筹),上海200437

出　　处：《药物生物技术》2010年第3期203-206,共4页Pharmaceutical Biotechnology

基　　金："重大新药创制"科技重大专项(2009ZX09301-007)

摘　　要：表柔红霉素是表阿霉素的半合成前体,通过中断dnmV基因,并将酮基还原酶aveBⅣ基因导入,可以将柔红霉素产生菌改造成表柔红霉素产生菌。在此基础上构建含有两个aveBⅣ表达单元的整合型质粒,并将其转入原有的表柔红霉素产生菌MHJ-02-30-1,使菌株内aveBⅣ的表达拷贝数由1个增加到3个。发酵验证表明,增加aveBⅣ基因拷贝数可以显著提高表柔红霉素的产量。目的菌株比出发菌株表柔红霉素的发酵单位提高了一倍以上。本文的工作为表柔红霉素发酵的产业化提供了有效途径。4＇-Epidaunorubicin currently is the main precursor in the semisynthesis of epidoxorubicin. Homotogous recombination was used for dnmV gene replacement and αveB Ⅳ integration in daunorubicin-producing strain to produce 4＇-epidaunorubin. Another integration plasmid containing two αveB Ⅳ expression modules was constructed and transformed into 4＇-epidaunorubin-producing strain MHJ-02-30-1. The copy-number of αveB Ⅳ was increased from one to three. The production of 4＇-epidaunorubicin was improved 2 fold by the increase of the expression modules of αveB Ⅳ compared with the original strain. The method described here provides the means to produce 4＇ epidaunorubicin efficiently on industrial scale.

关 键 词：表柔红霉素 基因置换 基因倍增 酮基还原酶 

分 类 号：R978.15[医药卫生—药品]