High-level Expression of Cecropin X in Escherichia coli

International Journal of Molecular Sciences

Yi Shen²  Xue Gang Lao¹  Yuan Chen²  Hong Zu Zhang² 

[1] Zhejiang Medicine Company, Ltd., Xinchang Pharmaceutical Factory, Xinchang County, Zhejiang Province 312500, P.R. China; E-mail:;State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, Nangjing University, Nanjing 210093, P.R. China

关键词: Cecropin X;    Escherichia coli;    Inclusion body;    Fermentation;    Tumorcidal activities;   

DOI  :  10.3390/i8060479

来源: mdpi

【 摘 要 】

Cecropin X is a short cationic peptide with a broad antibacterial and antitumor spectrum. Here, we report the production of a tumor necrosis factor (TNFα)-cecropin X fusion protein under the control of a temperature-inducible P_R promoter in the bacterial expression vector pRC. During fermentation, we studied and optimized essential parameters including the type of host cells, medium, timing of induction, post-induction time and dissolved oxygen level. Using the suitable conditions in the fermentation, up to 20 % ~ 23 % of the total cellular proteins is produced as the fusion protein, mostly in the form of inclusion bodies. After washing, on average about 5.27 g dried inclusion bodies could be collected from 1 L broth and the purity of inclusion bodies reached 80 %. Cecropin X obtained by cleaving the fusion protein with cyanogen bromide showed remarkable tumorcidal activity against mouse Lewis lung carcinoma 3LL in vivo.

【 授权许可】

Unknown   
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