重组大肠杆菌产疏绵状嗜热丝孢菌脂肪酶分批补料发酵工艺  

Optimization of fed-batch fermentation for Thermomyces lanuginosus lipase production with recombinant Eschericha coli

作　　者：马红叶[1] 郑仁朝[1] 赵川东[1] 郑裕国[1] 

机构地区：[1]浙江工业大学生物工程研究所 教育部生物转化与生物净化工程研究中心,浙江杭州310032

出　　处：《生物加工过程》2015年第2期9-12,18,共5页Chinese Journal of Bioprocess Engineering

基　　金：浙江省重大科技专项重大社会发展项目(2012C03005-2)

摘　　要：为了获得低成本的疏绵状嗜热丝孢菌脂肪酶(TLL),在5L发酵罐发酵中对工程菌E.coliBL21(DE3)/pET28b-TLL的分批补料发酵工艺进行研究。结果表明:以葡萄糖为碳源的补料培养基,采用溶氧(DO)反馈补料策略进行补料,当OD600=60时降温至28℃,分2次加入终质量浓度为30g/L乳糖进行诱导表达。优化后TLL的表达量提高到798.5U/L,是优化前的2.4倍。本研究为规模化发酵重组菌生产TLL奠定了基础。To establish cost-effective Thermomyces lanuginosus lipase（TLL） production process,fedbatch fermentation of the recombinant Eschericha coli BL21（DE3）/p ET28b-TLL was optimized in a 5-L bioreactor.The fed-batch medium with glucose as the main carbon source was fed by dissolved oxygen（DO） feedback method.The temperature was lowered down to 28 ℃ until the optical density at 600 nm reached about 60.And lactose was added twice at a final concentration of 30 g/L to induce expression.Under the optimal culture conditions,the volumetric activity of TLL reached 798.5 U/L that was 2.4-fold higher than that before the optimization.This findings will provide basis industrial production of TLL.

关 键 词：疏绵状嗜热丝孢菌脂肪酶 分批补料 发酵 大肠杆菌 

分 类 号：Q815[生物学—生物工程]