摘要
探索分泌型重组人胸腺素1α工程菌的高密度高表达发酵工艺.在摇瓶培养获得大肠杆菌分泌型重组人胸腺素1α融合蛋白发酵的最佳条件的基础上,用15 L自控发酵罐进行分批补料培养,发酵中分阶段限制性流加氮、碳源,保持溶氧在30%以上,OD600约为25的时候,添加0.5 mmol/LIPTG于28℃诱导10 h.最终菌体密度OD600可达40以上,目的产物占菌体总蛋白25%以上.分泌型人胸腺素1α基因工程菌高密度发酵工艺的建立为工业化生产重组人胸腺素1α提供基础.
To research the high cell density and high-expression fermentation of secretory recombinant human thymosin α1, the fed-batch fermentation was carried out in 15 L automatic fermentor on the basis of the optimized culture conditions obtained from the process of flask shaking culture. During the process of fermentation, nitrogen and carbohydrate were controlled at a special speed. The dissolved oxygen was maintained around 30%. And the induction started as OD600 reached about 25 by adding IPTG to a final concentration of 0.5 mmol/L and then continued incubation 10 hours at 28℃. The final cell density （OD600） exceeded 40, and the recombinant protein expression level was over 25% of the total cellular protein. The study provides a basis for largescale production of human thymosin α1 in industrial fermentation.
出处
《浙江工业大学学报》
CAS
2006年第3期277-280,共4页
Journal of Zhejiang University of Technology
基金
浙江省科技厅资助项目(No.2005C33031)
关键词
高密度发酵
胸腺素Α1
大肠杆菌
表达
high cell density fermentation
thymosin α1
E. coli
expression
