重组人肝细胞生长因子β链工程菌的高密度发酵

High cell density fermentation of recombinant human hepatocyte growth factor β chain bacteria

目的 实现重组人肝细胞生长因子 β链 (rhHGFβ)工程菌的高密度高表达发酵。 方法 首先在摇瓶中进行了培养条件的摸索 ,确定了该工程菌的培养条件、诱导起始时间和诱导时间 ,然后用 15L自控发酵罐进行分批补料培养 ,发酵中分阶段限制性流加氮、碳源 ,保持溶氧在 30 %以上。结果 菌体发酵密度达到 39g/L(湿菌重 ) ,达到并超过了重组蛋白在摇瓶中的表达水平 ,重组蛋白的表达占菌体总蛋白的 30 %左右。结论 本研究为rhHGFβ进一步下游纯化及功能研究奠定了基础。

Objective To research high cell density and high-expression fermentation of recombinant human hepatocyte growth factor β chain bacteria(DH5α-pBVhHGFβ). Methods Firstly,flask shaking culture was done to obtain optimized culture conditions including induction initiation time and induction lasting time. Then the fed-batch fermentation was carried out in 15 L automatic fermentor.During that time, nitrogen and carbohydrate were controlled at special speed.Dissolved oxygen was maintained around 30%. Results The final cell density was 39 g/L(wet cell weight). The expression was about 30% of total bacterial proteins. Conclusion The study provides a basis for further purification and function research.