重组人胰岛素样生长因子-Ⅰ工程菌高密度发酵工艺优化  

Optimization of high cell-density fermentation procedure of recombinant human insulin-like growth factor Ⅰ in E. coli

作　　者：陈蔚青[1] 陈虹[1] 胡文浪[1] 张建芬[1] 

机构地区：[1]浙江树人大学生物与环境工程学院,浙江杭州310015

出　　处：《中国生化药物杂志》2008年第2期121-124,共4页Chinese Journal of Biochemical Pharmaceutics

摘　　要：目的优化表达重组人胰岛素样生长因子-Ⅰ(IGF-Ⅰ)工程菌的高密度发酵条件。方法考察培养基以及诱导时机、诱导剂量和诱导时间对蛋白表达的影响;用自控发酵罐进行分批补料培养实验,确定优化工艺条件。结果以2×YT+0.5%葡萄糖为发酵培养基,经0.8 mmol/L IPTG(异丙基-β-D-硫代半乳糖苷)诱导5 h,通过pH-stat反馈补料方式实现工程菌高密度发酵与目的蛋白高效表达,每1 L发酵液收获干菌体50.1 g,IGF-Ⅰ含量达5.25 g/L。结论建立了IGF-Ⅰ工程菌优化的高密度发酵工艺,为IGF-Ⅰ的下游纯化和工业化生产奠定了基础。Purpose To investigate the optimal high cell-density fermentation procedure of recombinant human insulin-like growth factor Ⅰ （IGF- Ⅰ ） in E. coli. Methods 3 types of media, induction time and IPTG concentration have been analysed. To explore optimal fermentation conditions for expressing the recombinant protein, fed batch fermentation was carried out with autocontrol fermentor. Results Cultivated in 2 ×YT ＋ 0.5% glucose medium, induced by 0.8 mmol/L IPTG for 5 h, controlling dissolved oxygen and by pH-stat feeding solution, high eell-densityand high protein expression were achieved. Under the established conditions, 50.1 g dry cell weight （DCW）/L could be obtained, and 5.25 g/L IGF-Ⅰ was achieved. Conclusion The established high cell-density fermentative procedure has provided a useful base for further purification and large scale production of IGF-Ⅰ .

关 键 词：IGF—Ⅰ 工程菌 高密度发酵 

分 类 号：TQ920.6[轻工技术与工程—发酵工程]