Int. J. Mol. Sci. 2007, 8(6), 478-491; doi:10.3390/i8060479
Article

High-level Expression of Cecropin X in Escherichia coli

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
* 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
(This article belongs to the Special Issue Interaction of Biological Molecules)
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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.

AMA Style

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.

Chicago/Turabian Style

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.

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