摘要
目的建立重组人胰岛素样生长因子-1(hIGF-1)在大肠杆菌高密度发酵中表达的工艺。方法以工程菌DH5α/pET32a(IGF-1)为对象,通过分批发酵培养优化溶解氧浓度、pH及温度等发酵条件,在此基础上以甘油为限制性基质进行分批补料发酵,考察不同补料方式对发酵的影响。结果适合重组hIGF-1在工程菌高密度发酵中表达的条件:在菌株活化与初始培养后,诱导时控制发酵系统溶解氧浓度为20%,温度为30℃,pH为7.2并以恒定pH反馈补料,发酵12h菌体密度OD600达44.53,得菌体干质量17.49g/L,平均生产强度为1.458g.L-1.h-1,其中重组hIGF-1的相对表达量为32.5%。结论研究为工业化生产hIGF-1奠定了基础。
Objective To establish the expression process of recombinant human insulin-like growth factor-1 by high cell density fermentation of Escherichia coli. Method Taking engineering strain DH5a/pET32a（IGF-1） as object, the fermentation factors such as oxygen,pH and temperature were optimized through batch fermentation, then fed-batch fermentations were carried out using glycerol as carbon source so as to discuss the effects on fermentation by different feeding modes. Results The optimum conditions of recombined hlGF-1 expression in the engineering strain in high cell density fermentation were found to be. after activation and initial cultivation, the dissolved oxygen concentration was maintained at 20%, the temperature was adjusted to 30 ℃ ,and the culture pH was kept at 7.2 from induction, then pHstat feedback feeding-method was used for fed-batch fermentation. After 12 h, cell density OD600 of the engineering bacterial reached 44.53 with the dry cell weight of 17.49 g/L, the average production intensity was about 1. 458 g ·L^-1·h^-1 , and the relative expression level of hIGF-1 was about 32.5%. Conclusion The study provides a basis for large-scale production of hIGF-1 in industrial fermentation.
出处
《福建医科大学学报》
2013年第1期6-10,共5页
Journal of Fujian Medical University
基金
福建省高校产学合作科技重大项目(2010Y4002)
关键词
高密度发酵
重组大肠杆菌
人胰岛素样生长因子-1
high cell density fermentation
recombinant Escheriehia coli
human insulin-like growth factor-1
