关键词: 重组，遗传;大肠杆菌;发酵;肿瘤坏死因子;肽类

    摘 要:目的 探讨重组肿瘤导向多肽LTL-TNF工程菌的大规模培养与表达方法以获取重组LTLTNF. 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5L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DH5α/pBV-LTLTNF在5L发酵罐中培养至终密度A600nm 40～50时，目的蛋白的表达最高，可达菌体总蛋白的50%，LTL-TNF的含量达到5.12g·L-1 ，发酵总时间在12h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LTLTNF工程菌的工业化生产奠定了基础.

    Production of recombinant tumor targeting peptide LTL┐TNF by high cell density culture of Escherichia coli with DO┐stat fed┐batch condition

    YAN Zhen，ZHANG Ying-Qi，LI Bo，ZHAO Ning，SHI Ji-Hong，HAN Wei，WANG Li-Bing，WANG Zeng-Lu

    Biotechnology Center，Fourth Military Medical U-niversity，Xi'an710033，China

    Keywords:recombination，genetic;Escherichia.coli;fermen-tation，tumor necrosis factor;peptides

    Abstract:AIM High cell density cultivation research on re-combinant 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 on5L automatic fermentor.RESULTS By keeping dissolved oxygen at30%～40%and limiting glucose feeding during fermentation ......