类人胶原蛋白工程菌质粒稳定性研究

Plasmid instability of recombinant Escherichia coli in fed-batch fermentation

目的研究基因重组大肠杆菌补料分批发酵生产类人胶原蛋白过程中质粒稳定性。方法通过平板复制法,观察质粒稳定性在不同生长周期、卡那霉素浓度和不同比生长速率中的变化。结果质粒稳定性受生长周期、卡那霉素和比生长速率的影响。在μ为0.15、卡那霉素浓度为0.005g/L的情况下,工程菌生产效率最理想。结论通过控制发酵过程中的质粒稳定性可以有效地提高菌体产量和目标蛋白产量。

Aim To examine the plasmid instability of recombinant Escherichia coli during fed-batch fermentation. Methods The changes of the plasmid in stability under differnent conditions of growth stages, kanamycin and specific growth rate were observed with the methed of plate lysate. Results It showed that plasmid instability was influenced by the factors of growth stages, kanamycin concentration and specific growth rate. An effective goal was gained when the specific growth rate μ = 0. 15 and the kanamycin concentration was 0. 005 g/L. Conclusion The output of the biomass and protein could be effectively enhanced by the controllment of the plasmid instablity.