摘要
目的 探讨重组肿瘤导向多肽 L TL - TNF工程菌的大规模培养与表达方法以获取重组 L TL TNF.方法 首先在试管和三角瓶中探讨工程菌的生长和表达规律 ,筛选其最适宿主菌、最佳培养及诱导表达条件 ,然后在 5 L 自控发酵罐中进行分批补料培养 .结果 保持培养过程中 30 %～ 40 %的溶解氧和限制性流加葡萄糖 ,工程菌 DH5 α/ p BV- L TL TNF在5 L 发酵罐中培养至终密度 A6 0 0 nm 40～ 5 0时 ,目的蛋白的表达最高 ,可达菌体总蛋白的 5 0 % ,L TL - TNF的含量达到5 .12 g· L- 1 ,发酵总时间在 12 h左右 .结论 确定了周期短、产率高且稳定可靠的发酵工艺 ,为重组肿瘤导向多肽L TL TNF工程菌的工业化生产奠定了基础 .
AIM High cell density cultivation research on recombinant E.coli to produce recombinant tumor targeting peptide LTL TNF. METHODS To determine its optimized host cell, culture and induction condition, this research first focused on understanding the growth and expression specifici ty of recombinant LTLTNF on test tube and flask shaker. Then the fed batch culture was carried out on 5L automatic fermentor. RESULTS By keeping dissolved oxygen at 30%~40% and limiting glucose feeding during fermentation, the recombinant protein LTLTNF production of DH5 α/pBV LTLTNF in 5L fermentor reached 5.12 g·L -1 , about 50% of total amount of bacterial proteins, with the final cell density of A 600nm 40~50 in around 12 h. CONCLUSION A short cycle, high expression and stabilized fermentation process of recombinant LTLTNF has been established.
出处
《第四军医大学学报》
北大核心
2001年第14期1297-1300,共4页
Journal of the Fourth Military Medical University
