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 Okano, K.
Right arrow Articles by Kondo, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Okano, K.
Right arrow Articles by Kondo, A.
Agricola
Right arrow Articles by Okano, K.
Right arrow Articles by Kondo, A.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, February 2008, p. 1117-1123, Vol. 74, No. 4
0099-2240/08/$08.00+0     doi:10.1128/AEM.02012-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

System Using Tandem Repeats of the cA Peptidoglycan-Binding Domain from Lactococcus lactis for Display of both N- and C-Terminal Fusions on Cell Surfaces of Lactic Acid Bacteria{triangledown}

Kenji Okano,1 Qiao Zhang,2 Sakurako Kimura,3 Junya Narita,1 Tsutomu Tanaka,4 Hideki Fukuda,4 and Akihiko Kondo2*

Department of Molecular Science and Material Engineering, Graduate School of Science and Technology,1 Department of Chemical Science and Engineering, Graduate School of Engineering,2 Department of Chemical Science and Engineering, Faculty of Engineering,3 Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe 657-8501, Japan4

Received 3 September 2007/ Accepted 10 December 2007

Here, we established a system for displaying heterologous protein to the C terminus of the peptidoglycan-binding domain (cA domain) of AcmA (a major autolysin from Lactococcus lactis). Western blot and flow cytometric analyses revealed that the fusion proteins (cA-AmyA) of the cA domain and {alpha}-amylase from Streptococcus bovis 148 (AmyA) are efficiently expressed and successfully displayed on the surfaces of L. lactis cells. AmyA was also displayed on the cell surface while retaining its activity. Moreover, with an increase in the number of cA domains, the quantity of cA-AmyA fusion proteins displayed on the cell surface increased. When three repeats of the cA domain were used as an anchor protein, 82% of {alpha}-amylase activity was detected on the cells. The raw starch-degrading activity of AmyA was significantly higher when AmyA was fused to the C terminus of the cA domain than when it was fused to the N terminus. In addition, cA-AmyA fusion proteins were successfully displayed on the cell surfaces of Lactobacillus plantarum and Lactobacillus casei.

* Corresponding author. Mailing address: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe 657-8501, Japan. Phone and fax: 81-78-803-6196. E-mail: akondo{at}kobe-u.ac.jp

{triangledown} Published ahead of print on 21 December 2007.

Applied and Environmental Microbiology, February 2008, p. 1117-1123, Vol. 74, No. 4
0099-2240/08/$08.00+0     doi:10.1128/AEM.02012-07
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»