重组人锰超氧化物歧化酶高密度发酵培养条件研究

目的:研究发酵罐中采用乳糖诱导高效表达重组人锰超氧化物歧化酶(rhMn-SOD)的条件。方法:通过摇瓶和发酵罐培养研究了培养基、乳糖浓度、诱导时机、表达时间、溶氧条件、补料碳源种类对rhMn-SOD表达的影响。结果:乳糖诱导浓度为0.25%,溶氧浓度为40%～60%,用葡萄糖作为补料碳源进行高密度发酵,菌体密度A600达到28.98,菌体湿重达到60g/L,Mn-SOD活性达到18222.22U/g,比活为1057.5U/mg,实现了rhMn-SOD的高效表达。结论:对培养基配方和发酵条件进行了优化,为rhMn-SOD大规模生产奠定了基础。

Optimization of high-cell-density fermentation condition of recombinant human manganese superoxide dismutase

Aim:To investigate the fermertation condition of lactose-induced recombinant human manganese superoxide dismutase(rhMn-SOD) in the bioreactor.Methods:The influences of culture medias,lactose concentration,inducing times,expression time,dissolved oxygen level and carbon source on rhMn-SOD expression were studyed both in the shake falsks and the bioreactor.Results:In the high-cell-density fermentation,after the bacteria being induced by 0.25% lactose,took glucose as carbon source with the dissolved oxygen controlled between 40% and 60%,the optical-density(A600) of bacteria reached 29.98,and the wet cell weight got 60 g/L.Furthermore,Mn-SOD activity reached 18 222.22 U/g and the specific activity of Mn-SOD was 1 057.5 U/mL.Conclusion:Efficient expression of rhMn-SOD in E.coli was achieved by the optimized fermentation conditions which laid the foundation for its mass production.