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High-level Expression of Cecropin X in Escherichia coli
State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, Nangjing University, Nanjing 210093, P.R. China
Zhejiang Medicine Company, Ltd., Xinchang Pharmaceutical Factory, Xinchang County, Zhejiang Province 312500, P.R. China
* Author to whom correspondence should be addressed.
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 antitumorspectrum. Here, we report the production of a tumor necrosis factor (TNFα)-cecropin Xfusion protein under the control of a temperature-inducible PR promoter in the bacterialexpression vector pRC. During fermentation, we studied and optimized essentialparameters including the type of host cells, medium, timing of induction, post-inductiontime and dissolved oxygen level. Using the suitable conditions in the fermentation, up to20 % ~ 23 % of the total cellular proteins is produced as the fusion protein, mostly in theform of inclusion bodies. After washing, on average about 5.27 g dried inclusion bodiescould 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 showedremarkable tumorcidal activity against mouse Lewis lung carcinoma 3LL in vivo.
Keywords: Cecropin X; Escherichia coli; Inclusion body; Fermentation; Tumorcidal activities
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MDPI and ACS Style
Shen, Y.; Lao, X.G.; Chen, Y.; Zhang, H.Z.; Xu, X.X. High-level Expression of Cecropin X in Escherichia coli. Int. J. Mol. Sci. 2007, 8, 478-491.
Shen Y, Lao XG, Chen Y, Zhang HZ, Xu XX. High-level Expression of Cecropin X in Escherichia coli. International Journal of Molecular Sciences. 2007; 8(6):478-491.
Shen, Yi; Lao, Xue G.; Chen, Yuan; Zhang, Hong Z.; Xu, Xian X. 2007. "High-level Expression of Cecropin X in Escherichia coli." Int. J. Mol. Sci. 8, no. 6: 478-491.