高密度培养大肠杆菌TB1pMALOCIFm和高表达重组融合人破骨细胞形成抑制因子成熟肽研究.pdfVIP

高密度培养大肠杆菌TB1/pMAL—OCIFm和高表达重组融合人破骨细胞形成抑制因 子成熟肽 1 2 2 1 张兴群 王梁华 焦炳华 袁勤生 1 华东理工大学生物反应器工程国家重点实验室 上海 200237 2 第二军医大学基础部生化与分子生物学教研室 上海 200433 摘 要 目的 高密度 高表达培养重组大肠杆菌TB1/pMAL-hOCIFm生产重组人破骨细胞 形成抑制因子(rhOCIFm)方法和结果 应用Bioengerring3.7L自控发酵罐 采用分 批培养和补料分批培养相结合的培养技术 控制碳源和氮源的补加 控制溶解氧 使重组菌 E. coli TB1/pMAL— hOCIFm发酵菌体OD600达到58.6人重组破骨细胞形成抑制因子MBP 融合蛋白(MBP-hOCIFm)的含量达到3.2 g/L结论 本研究为工业化生产重组 hOCIFm奠定 了基础 关键词 破骨细胞形成抑制因子(OCIF)表达 高密度培养 High cell-density culture of E.coli TB1/pMAL OCIFm and high expression of recombinant human Osteoclastogenesis Inhibitory factor(OCIF Mature peptide 1 2 2 1 Zhang Xing-Qun , Wang liang-Hua , Jiao Bing-Hua , Yuan Qin-Sheng and Technolo of Biochemistry and Molecular Biology,Department of Basic Medicine,Second Military Medical University,Shanghai 200433,China) Abstract Objective High cell-density and high expression culture of E.coli TB1/pMAL-hOCIFm to produce recombinant human osteoclastogenesis inhibitory factor active domain(OCIFm). Methods and Results Batch and fed-batch culture process were used with the carbon-nitrogen control. DO-stat method in 3.7 L Bioengineering autocontrol fermentor was carried out to produce recombinant hOCIFm in E.coli TB1/pMAL- hOCIFm .The final cell density and concentration of recombinant hOCIFm fusion protein were 46.3D600 and 3.2 g/L. The purity of OCIFm fusion protein was more then 70% after affinity chromatography on Amylose Resin. Conclusion This study provides a basic work for production recombinant OCIFm in industrial scale. Key Words OCIF; fermentation; Fed-batch; expression