This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Tanaka, T.
Right arrow Articles by Matsunaga, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tanaka, T.
Right arrow Articles by Matsunaga, T.
Agricola
Right arrow Articles by Tanaka, T.
Right arrow Articles by Matsunaga, T.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, December 2008, p. 7600-7606, Vol. 74, No. 24
0099-2240/08/$08.00+0     doi:10.1128/AEM.00162-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Novel Method for Selection of Antimicrobial Peptides from a Phage Display Library by Use of Bacterial Magnetic Particles{triangledown}

Tsuyoshi Tanaka, Yoriko Kokuryu, and Tadashi Matsunaga*

Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16 Naka-Cho, Koganei, Tokyo 184-8588, Japan

Received 17 January 2008/ Accepted 16 October 2008

Antimicrobial peptides were isolated from a phage display peptide library using bacterial magnetic particles (BacMPs) as a solid support. The BacMPs obtained from "Magnetospirillum magneticum" strain AMB-1 consist of pure magnetite (50 to 100 nm in size) and are covered with a lipid bilayer membrane derived from the invagination of the inner membrane. BacMPs are easily purified from a culture of magnetotactic bacteria by magnetic separation. Approximately 4 x 1010 PFU of the library phage (complexity, 2.7 x 109) was reacted with BacMPs. The elution of bound phages from BacMPs was performed by disrupting its membrane with phospholipase D treatment. Six candidate peptides, which were highly cationic and could bind onto the BacMP membrane, were obtained. They exhibited antimicrobial activity against Bacillus subtilis but not against Escherichia coli and Saccharomyces cerevisiae. The amino acid substitution of the selected peptide, KPQQHNRPLRHK (peptide 6-7), to enhance the hydrophobicity resulted in obvious antimicrobial activity against all test microorganisms. The present study shows for the first time that a magnetic selection of antimicrobial peptides from the phage display peptide library was successfully achieved by targeting the actual bacterial inner membrane. This BacMP-based method could be a promising approach for a high-throughput screening of antimicrobial peptides targeting a wide range of species.

* Corresponding author. Mailing address: Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16 Naka-Cho, Koganei, Tokyo 184-8588, Japan. Phone: 81-42-388-7020. Fax: 81-42-385-7713. E-mail: tmatsuna{at}cc.tuat.ac.jp

{triangledown} Published ahead of print on 24 October 2008.

Applied and Environmental Microbiology, December 2008, p. 7600-7606, Vol. 74, No. 24
0099-2240/08/$08.00+0     doi:10.1128/AEM.00162-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»