粒细胞集落刺激因子_

用重组大肠杆菌生产rhGCSF高密度发酵的方法

http://www.100md.com 《第四军医大学学报》 2004年第12期

粒细胞集落刺激因子,,粒细胞集落刺激因子，重组；发酵；流加培养,0引言,1材料和方法,2结果,3讨论,【参考文献】

     A method of high density fermentation of Escherichia coli in producing rhGCSF

    WANG HaiBo, WANG LiLi, OU JunJie, GENG XinDu

    1Department of Chemistry, School of Basic Medicine, Fourth Military Medical University, Xian 710033, China, 2Institute of Modern Separation Science, Key Laboratory of Modern Separation Science in Shaanxi Province, Northwest University, Xian 710069, China

    【Abstract】 AIM: To optimize the production techniques of (recombinant human granulocyte colonystimulating factor, rhGCSF) and offer some theoretical basis for largescale production of rhGCSF in industrial fermentation. METHODS: Based on the experimental results obtained in shakeflask cultivation, the bacterial growth curves of recombinant E.coli were measured and the influence of culture pH, dissolved oxygen (DO) and inducing time at various levels on the growth of recombinant E. coli were investigated respectively in a 5 L bioreactor. The fedbatch cultivation conditions for the production of the target protein (rhGCSF) in engineered bacteria were studied and the stable fermentation parameters for higher yield of rhGCSF in the 5 L bioreactor were determined. RESULTS: The optimum fedbatch cultivation conditions were found to be: at the beginning of the fedbatch cultivation, the initial culture pH was adjusted to 7.2 by automated addition of 2 mol/L sodium hydroxide (NaOH) and at the time of expressing, the culture pH was kept at 6.8. The dissolved oxygen concentration was maintained at over 50% throughout the cultivation. The engineered bacteria were induced in the middle logarithm phase (A600 nm 12.0). Under these optimum conditions, the cell optical density (A600 nm) was (21.3±0.9), the dry cell weight (DCW) and the wet cell weight (WCW) reached (12.3±0.7) g/L and (43.8±0.9) g/L, respectively, and the expression of rhGCSF was (41.9±1.6)%. CONCLUSION: The growth of engineered bacteria and the expression of rhGCSF are influenced greatly by different cultivation conditions such as pH, dissolved oxygen and inducing time. ......

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