重组人瘦素大肠杆菌的高密度发酵与瘦素的纯化

High Density Fermentation of pET32a-OB[BL21(DE3)]and Purification of Human Leptin

[目的]探索获得大量高纯度重组人瘦素的方法。[方法]通过5 L发酵罐对重组人瘦素大肠杆菌pET32a-OB[BL21(DE3)]进行了高密度发酵,并通过DEAE sepharose Fast Flow阴离子层析分离将IPTG诱导表达的人瘦素进行纯化,并通过ESI-Q-TOF-MS/MS分析验证目的蛋白。[结果]得到的培养物为3 L左右,OD600为20.25,纯化后的人瘦素纯度为98%左右。[结论]发酵罐发酵得到的菌体密度要远大于正常用三角瓶培养的菌体密度,阴离子交换层析法制备重组人瘦素符合大量生产高纯度重组人瘦素的要求。

[Objective] The aim of this study was to discuss a method of obtaining mass of high-purity recombinant leptin. [Method]Highdensity fermentation of pET32a-OB[BL21（ DE3） ]by 5 L fermentation tank was conducted and human leptin expressed by IPTG induction waspurified with DEAE sepharose Fast Flow. And the target protein was identified by ESI-Q-TOF-MS/MS. [Result]The obtained product of fer-mentation was about 3 L and OD600 was 20. 25. The purity of purified human leptin was about 98% . [Conclusion]The density of medium ob-tained from fermentation by 5 L fermentation tank was greatly more than that by normal culture by triangular flask. The preparation of the re-combinant human leptin by DEAE sepharose Fast Flow could meet the demands of mass production of high-density recombinant leptin.