重组人血管生长素基因工程菌发酵条件的初步研究

Preliminary studies on fermentation of the recombinant human angiogenin in E.coli

目的研究表达重组人血管生长素 (rhANG)工程菌的发酵条件。方法采用摇瓶发酵 ,研究不同宿主菌对表达rhANG的影响 ,同时对培养基、诱导时期、诱导时间和初始pH等发酵条件以及质粒稳定性进行研究。结果选用E .coliCDH为宿主菌 ,在SOB培养基中培养至A6 0 0nm 为 0 .5～ 0 .6时 ,诱导表达 5h ,rhANG表达量最高达 30 %。重组质粒具有良好的分离稳定性和结构稳定性。结论此发酵条件可以较好地提高rhANG的表达量 。

PurposeTo study the fermentation process of recombinant bacteria E.coli CDH expressing recombinant human angiogenin.MethodsThe E.coli host cells, culture medium, induction period, induction time and pH were optimized. The stability of pBV220 hANG plasmid was also investigated. ResultsThe expression level of rhANG was about 30% of the total bacteria protein, when recombinant E.coli CDH harboring pBV220 hANG plasmid was induced for 5 hours after the bacteria density A 600nm reached 0.5～0.6 in SOB medium. The pBV220 hANG plasmid in E.coli CDH remained stable, after being cultured to 100 generations.ConclusionThe developed fermentation technology might increase the expression of rhANG, and is suitable for the fermentation of rhANG on a large scale.