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Applied and Environmental Microbiology, June 2009, p. 3980-3986, Vol. 75, No. 12
0099-2240/09/$08.00+0     doi:10.1128/AEM.02753-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Direct Expression of Antimicrobial Peptides in an Intact Form by a Translationally Coupled Two-Cistron Expression System{triangledown} ,{dagger}

Su A Jang,1 Bong Hyun Sung,1 Ju Hyun Cho,2 and Sun Chang Kim1*

Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 305-701, Korea,1 Department of Biology, Research Institute of Life Science, Gyeongsang National University, Jinju 660-701, Korea2

Received 3 December 2008/ Accepted 31 March 2009

We describe a novel prokaryotic expression system for the production of cationic antimicrobial peptides (AMPs). The method relies on a translationally coupled two-cistron system, in which the termination codon for the first cistron (which encodes the anionic polypeptide mIFc2, a derivative of human gamma interferon) overlaps with the initiation codon for the second cistron (which encodes a cationic AMP) in the sequence of 5'-TAATG-3'. By forming an insoluble complex with the AMP upon translation, the mIFc2 protein efficiently neutralized the toxicity of the coexpressed cationic AMP and minimized the sensitivity of AMP to proteolytic degradation in a host. The AMPs were retrieved from the insoluble inclusion bodies without any chemical or enzymatic cleavage step by simple cation-exchange chromatography. With our system, ~100 mg of various AMPs (buforin IIb, parasin I, and pexiganan) were obtained from 1 liter of Escherichia coli culture. Our expression system may represent a universal cost-effective solution for the mass production of intact AMPs in their natural forms.

* Corresponding author. Mailing address: Department of Biological Sciences, KAIST, 335 Gwahangno, Yuseong-gu, Daejeon 305-701, Korea. Phone: 82-42-350-2619. Fax: 82-42-350-2610. E-mail: sunkim{at}kaist.ac.kr

{triangledown} Published ahead of print on 10 April 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, June 2009, p. 3980-3986, Vol. 75, No. 12
0099-2240/09/$08.00+0     doi:10.1128/AEM.02753-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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