高密度培养大肠杆菌TB1/pMAL-hOCIFm的优化条件

High Cell-Density Culture of E.coli TB1/pMAL-hOCIFm

利用Bioengeering3.7L自控式发酵罐,以分批培养和补料分批培养相结合的培养技术,高密度培养重组大肠杆菌TB1/pMAL-hOCIFm,生产重组人破骨细胞形成抑制因子成熟肽(recombinanthumanosteoclastogenesisinhibitoryfactormaturepeptide,rhOCIFm)。通过对碳、氮源补加方式以及溶解氧等参数的控制,使工程菌E.coliTB1/pMAL-hOCIFm的发酵菌体OD600达到57.8,rhOCIFm与MBP融合蛋白(hOCIFm-MBP)的含量达到3.2g/L。本研究为工业化生产rhOCIFm奠定了基础。

Batch and fed-batch culture process of E. coli TB1/pMAL-hOCIFm were carried out to produce recombinant human osteoclastogenesis inhibitory factor mature peptide (rhOCIFm) in 3.7 L Bioengineering autocontrol fermentor. High cell-density and high expression were achieved by the control of carbon/nitrogen source and dissolved oxygen. The final cell density and concentration of rhOCIFm-MBP fusion protein were 57.8 OD600 and 3.2 g/L respectively. The purity of rhOCIFm-MBP fusin protein was more than 70% after treated with affinity chromatography on amylose resin. This study provides a basic work for the production of rhOCIFm in industrial scale.