A study of high cell density cultivation process of recombinant Helicobacter pylori multi-epitope vaccine engineering bacteria

International Journal of Clinical and Experimental Medicine

Xing Pan¹  Mingyuan Li¹  Baoning Wang¹  Lizhen Gao¹  Jie Zhu¹  Wanyi Li¹  Jing Yang¹  Hongren Wang¹  Yongjun Zhou¹ 

关键词: Helicobacter pylori;    Hp;    multi-epitope vaccine;    engineering bacteria;    high cell density cultivation;    HCDC;   

DOI  :  

学科分类：医学（综合）

来源: e-Century Publishing Corporation

【 摘 要 】

Objective: To establish high cell density cultivation process of recombinant Helicobacter pylori multi-epitope vaccine engineering bacteria BIB. Methods: Based on the results of shake flask fermentation, the process was magnified into volume of a 50 L fermenter to optimize and verify the factors affecting the yield of the target protein, such as the fermentation medium, working seed inoculation amount, inducer concentration, induction starting time, induction duration, inducer adding mode and feeding strategy. Results: After activated in modified TB medium at 37°C for 8 h, the BIB working seed was inoculated at 5% (v/v) and was induced for expression for another 11 h by the final concentration of 5 mmol/L lactose. In growth phase, glucose at rate of 80 ml/h was used as carbon source, and in induction phase, glycerol at rate of 40 ml/h was used as carbon source; ammonia water was added dropwise to control pH at about 7.0, and revolution speed is adjusted to control the dissolved oxygen at above 30%; ultimately the output of bacterial body was 70 g/L and protein expression amount was about 32%. Conclusion: After high cell density cultivation of the recombinant engineering bacteria, expression and yield of the target protein rBIB significantly increased.

【 授权许可】

Unknown