High-level Expression of Cecropin strongXstrong in Escherichia coli.pdfVIP

Int. J. Mol. Sci. 2007, 8, 478-491 International Journal of Molecular Sciences ISSN 1422-0067 © 2007 by MDPI /ijms/ Full Research Paper High-level Expression of Cecropin X in Escherichia coli Yi Shen 1,*, Xue Gang Lao 2, Yuan Chen 1, Hong Zu Zhang 1 and Xian Xiu Xu 1 1 State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, Nangjing University, Nanjing 210093, P.R. China 2 Zhejiang Medicine Company, Ltd., Xinchang Pharmaceutical Factory, Xinchang County, Zhejiang Province 312500, P.R. China; E-mail: laoxg@ * Author to whom correspondence should be addressed; Tel.: +86 025 Fax: +86 025 E-mails: shenyi@, hzzhang@ Received: 6 February 2007 / in Revised Form: 23 April 2007 / Accepted: 25 May 2007 / Published: 4 June 2007 Abstract: 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 PR 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, most