抗阿尔茨海默病神经保护肽[Gly14]-Humanin融合基因工程菌发酵工艺研究

High density fermentation procedure of recombinant E. coli strain expressing an anti-Alzheimer's disease neuroprotective peptide [Gly14]-Humanin fusion gene

目的优化大肠杆菌表达重组[Gly14]-Humanin(HNG)融合基因工程菌的发酵工艺。方法在15L发酵罐内,研究培养基、培养条件和诱导时间对工程菌生长和目的蛋白表达的影响,并考察工程菌中重组质粒的遗传稳定性。结果工程菌在pH7.4的2×YT培养基中诱导5h,菌体湿重可达80g/L,目的蛋白表达量约占总蛋白的36%,所构建的重组质粒在BL21宿主菌中传代稳定。结论优化了HNG融合基因工程菌的发酵和表达条件,为药物中试研究和规模化生产奠定了基础。

Objective To optimize the fermentation procedure of recombinant E. coli strain expressing [Gly14]- Humanin fusion gene. Methods We studied the influence of culture medium and durations for incubation and induction on the growth of recombinant E. coli and expression of target protein in a 15 L fermentor, and explored the genetic stability of recombinant plasmid. Results After the recombinant E. coli strain cultured in 2 x YT medium （pH 7.4） was induced for 5 h, the wet weight of bacteria reached 80 g/L. The expressed product contained about 36% of total somatic protein. The constructed recombinant plasmid was inherited steadily in host bacterial strain E. coli BL21. Conclusion The condition for fermentation of recombinant E. coli strain and expression of HNG fusion gene was optimized, which forms a basis for pilot drug study and massive production of HNG protein.