可溶型非融合血管生长抑制因子Kringle 5基因工程菌发酵培养和诱导表达条件的优化  

Optimization of Fermentation Medium and Induction Expression Conditions for A Genetic Engineering Bacterium Producing Soluble and Nonfusion Angiogenesis Inhibitor Kringle 5

作　　者：马晓娟[1] 蔚萍[1] 妙亮[1] 边六交[1] 

机构地区：[1]西北大学生命科学学院,陕西西安710069

出　　处：《化学与生物工程》2012年第4期27-31,共5页Chemistry & Bioengineering

基　　金：国家重大新药创制科技重大专项项目(2009ZX09103-696)

摘　　要：在实验室试管培养条件下优化了基因工程菌E.coli BL21(DE3)pET15b/K5发酵产可溶型非融合血管生长抑制因子Kringle 5的培养基和诱导表达条件。经菌体干重测定和SDS-PAGE检测,确定最佳培养基(g.L-1)为:胰蛋白胨10.0,酵母提取物5.0,NaCl 10.0,葡萄糖6.0,NH4Cl 2.6,NaH2PO45.0,Na2HPO46.0;优化的诱导表达条件为:诱导剂浓度0.01mmol.L-1,诱导时间6h,诱导温度37℃,摇床转速220r.min-1。在优化的培养基和诱导表达条件下,菌体干重为1.8g.L-1,Kringle 5表达量为360mg.L-1,占总蛋白含量的20%,与基础LB培养基相比,Krin-gle 5表达量提高了1.18倍。The optimization of fermentation medium and induction expression conditions for genetic engineering bacterium E.coli BL21（DE3） pET15b/K5 producing soluble and nonfusion angiogenesis inhibitor Kringle 5 were investigated.The results showed that the optimum fermentation medium（g·L-1） was：tryptone 10.0,yeast extract 5.0,NaCl 10.0,glucose 6.0,NH4Cl 2.6,NaH2PO4 5.0,Na2HPO4 6.0,and the optimum induction expression conditions were：IPTG concentration 0.01 mmol·L-1,induction time 6 h,induction temperature 37 ℃,rotational speed 220 r·min-1.Under the optimum fermentation medium and induction expression conditions,the cell dry weight was 1.8 g·L-1,the expression amount of Kringle 5 was 360 mg·L-1,which occupied 20% of total bacterial protein and increased 2.18-fold as compared with LB medium.

关 键 词：血管生长抑制因子Kringle5 发酵培养基 诱导表达 基因工程菌 优化 

分 类 号：TQ464[化学工程—制药化工] Q789[生物学—分子生物学]