重组人血管内皮细胞生长因子工程菌高密度发酵工艺研究  

High cell density fed-batch culture of E.coli expressing rhVEGF121

作　　者：胡志明[1] 马骊[1] 周明乾[1] 孟民杰[1] 杨介钻[1] 王小宁[1] 

机构地区：[1]南方医科大学分子免疫学研究所,广东广州510515

出　　处：《第一军医大学学报》2005年第3期267-269,共3页Journal of First Military Medical University

基　　金：广州市科技计划(2003Z2-E0171);广东省科技攻关计划(2004B31201010);军队医药卫生科研基金(04LX028)~~

摘　　要：目的建立人血管内皮细胞生长因子(BL21/pET-24a/hVEGF121)工程菌的高密度发酵工艺。方法采用发酵罐发酵,对影响工程菌生长及目的蛋白表达的因素如培养基、诱导时间及补料进行优化。结果采用M9复合培养基、活化至对数生长期时诱导4 h以及以甘油为碳源连续流加补料的条件发酵,可使菌体量提高至68 g/L,rhVEGF121的表达量达菌体蛋白总量的23%。结论该发酵工艺提高了工程菌的产量和rhVEGF121的表达水平。Objective To investigate the optimal high cell density fermentation conditions of recombinant E.coli BL21/pET- 24a/hVEGF121 expressing recombinant human vascular endothelial growth factor (rhVEGF121). Methods The effects of the composition of the fermentation medium, induction time and fed-batch carbon sources on the expression level of rhVEGF121 and cell output were analyzed. Results and Conclusion When cultured in modified M9 medium and induced for 4 h in the presence of 0.5 mmol/L IPTG at 37 ℃ with glycerol as the carbon sources by continuous fed-batch mode, the recombinant E.coli expressed rhVEGF121 at the level up to 23% of the total proteins and the yield reached 68 g/L. The optimized fermentation condition for recombinant E.coli enables high expression level of rhVEGF121.

关 键 词：人血管内皮细胞生长因子 高密度发酵 大肠杆菌 

分 类 号：TQ920[轻工技术与工程—发酵工程]