| 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 |
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| 引用本文： | 胡志明,马骊,周明乾,孟民杰,杨介钻,王小宁.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[J].第一军医大学学报,2005,25(3):267-269. |
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| 作者姓名： | 胡志明 马骊 周明乾 孟民杰 杨介钻 王小宁 |
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| 作者单位： | 南方医科大学分子免疫学研究所,广东广州510515 |
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| 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.
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| 关 键 词： | 人血管内皮细胞生长因子 高密度发酵 大肠杆菌 |
High cell density fed-batch culture of E.coli expressing rhVEGF121] |
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| Zhi-Ming Hu,Li Ma,Ming-Qian Zhou,Min-Jie Meng,Jie-Zuan Yang,Xiao-Ning Wang.High cell density fed-batch culture of E.coli expressing rhVEGF121][J].Journal of First Military Medical University,2005,25(3):267-269. |
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| Authors: | Zhi-Ming Hu Li Ma Ming-Qian Zhou Min-Jie Meng Jie-Zuan Yang Xiao-Ning Wang |
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| Institution: | Institute of Molecular Immunology, Southern Medical University, Guangzhou 510515, China. hzm@fimmu.com |
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| Abstract: | OBJECTIVE: To investigate the optimal high cell density fermentation conditions of recombinant E.coli BL21/pET- 24a/hVEGF(121) expressing recombinant human vascular endothelial growth factor (rhVEGF(121)). METHODS: The effects of the composition of the fermentation medium, induction time and fed-batch carbon sources on the expression level of rhVEGF(121) 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 degrees celsius; with glycerol as the carbon sources by continuous fed-batch mode, the recombinant E.coli expressed rhVEGF(121) 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 rhVEGF(121). |
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