This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow An author's correction has been published
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 HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Turner, M. S.
Right arrow Articles by Giffard, P. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Turner, M. S.
Right arrow Articles by Giffard, P. M.
Agricola
Right arrow Articles by Turner, M. S.
Right arrow Articles by Giffard, P. M.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, October 2003, p. 5855-5863, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.5855-5863.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Peptide Surface Display and Secretion Using Two LPXTG-Containing Surface Proteins from Lactobacillus fermentum BR11

Mark S. Turner,* Louise M. Hafner, Terry Walsh, and Philip M. Giffard

Infectious Diseases Program, Faculty of Science, Queensland University of Technology, Brisbane, Australia

Received 4 March 2003/ Accepted 9 July 2003

A locus encoding two repetitive proteins that have LPXTG cell wall anchoring signals from Lactobacillus fermentum BR11 has been identified by using an antiserum raised against whole L. fermentum BR11 cells. The first protein, Rlp, is similar to the Rib surface protein from Streptococcus agalactiae, while the other protein, Mlp, is similar to the mucus binding protein Mub from Lactobacillus reuteri. It was shown that multiple copies of mlp exist in the genome of L. fermentum BR11. Regions of Rlp, Mlp, and the previously characterized surface protein BspA were used to surface display or secrete heterologous peptides in L. fermentum. The peptides tested were 10 amino acids of the human cystic fibrosis transmembrane regulator protein and a six-histidine epitope (His6). The BspA promoter and secretion signal were used in combination with the Rlp cell wall sorting signal to express, export, and covalently anchor the heterologous peptides to the cell wall. Detection of the cell surface protein fusions revealed that Rlp was a significantly better surface display vector than BspA despite having lower cellular levels (0.7 mg per liter for the Rlp fusion compared with 4 mg per liter for the BspA fusion). The mlp promoter and encoded secretion signal were used to express and export large (328-kDa at 10 mg per liter) and small (27-kDa at 0.06 mg per liter) amino-terminal fragments of the Mlp protein fused to the His6 and CFTR peptides or His6 peptide, respectively. Therefore, these newly described proteins from L. fermentum BR11 have potential as protein production and targeting vectors.

* Corresponding author. Mailing address: Infectious Diseases Program, Faculty of Science, Queensland University of Technology, G.P.O. Box 2434, Brisbane, Queensland 4001, Australia. Phone: (61-7) 3864-2567. Fax: (61-7) 3864-1534. E-mail: ms.turner{at}qut.edu.au.

Applied and Environmental Microbiology, October 2003, p. 5855-5863, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.5855-5863.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Lo, R., Turner, M. S., Barry, D. G., Sreekumar, R., Walsh, T. P., Giffard, P. M. (2009). Cystathionine {gamma}-Lyase Is a Component of Cystine-Mediated Oxidative Defense in Lactobacillus reuteri BR11. J. Bacteriol. 191: 1827-1837 [Abstract] [Full Text]  
  • Tan, Y. P., Giffard, P. M., Barry, D. G., Huston, W. M., Turner, M. S. (2008). Random Mutagenesis Identifies Novel Genes Involved in the Secretion of Antimicrobial, Cell Wall-Lytic Enzymes by Lactococcus lactis. Appl. Environ. Microbiol. 74: 7490-7496 [Abstract] [Full Text]  
  • Bolduc, G. R., Madoff, L. C. (2007). The group B streptococcal alpha C protein binds {alpha}1 1-integrin through a novel KTD motif that promotes internalization of GBS within human epithelial cells. Microbiology 153: 4039-4049 [Abstract] [Full Text]  
  • Turner, M. S., Lo, R., Giffard, P. M. (2007). Inhibition of Staphylococcus aureus Growth on Tellurite-Containing Media by Lactobacillus reuteri Is Dependent on CyuC and Thiol Production. Appl. Environ. Microbiol. 73: 1005-1009 [Abstract] [Full Text]  
  • Narita, J., Okano, K., Kitao, T., Ishida, S., Sewaki, T., Sung, M.-H., Fukuda, H., Kondo, A. (2006). Display of {alpha}-Amylase on the Surface of Lactobacillus casei Cells by Use of the PgsA Anchor Protein, and Production of Lactic Acid from Starch. Appl. Environ. Microbiol. 72: 269-275 [Abstract] [Full Text]  
  • Buck, B. L., Altermann, E., Svingerud, T., Klaenhammer, T. R. (2005). Functional Analysis of Putative Adhesion Factors in Lactobacillus acidophilus NCFM. Appl. Environ. Microbiol. 71: 8344-8351 [Abstract] [Full Text]  
  • Pretzer, G., Snel, J., Molenaar, D., Wiersma, A., Bron, P. A., Lambert, J., de Vos, W. M., van der Meer, R., Smits, M. A., Kleerebezem, M. (2005). Biodiversity-Based Identification and Functional Characterization of the Mannose-Specific Adhesin of Lactobacillus plantarum. J. Bacteriol. 187: 6128-6136 [Abstract] [Full Text]  
  • Walter, J., Chagnaud, P., Tannock, G. W., Loach, D. M., Dal Bello, F., Jenkinson, H. F., Hammes, W. P., Hertel, C. (2005). A High-Molecular-Mass Surface Protein (Lsp) and Methionine Sulfoxide Reductase B (MsrB) Contribute to the Ecological Performance of Lactobacillus reuteri in the Murine Gut. Appl. Environ. Microbiol. 71: 979-986 [Abstract] [Full Text]  
  • Lindahl, G., Stalhammar-Carlemalm, M., Areschoug, T. (2005). Surface Proteins of Streptococcus agalactiae and Related Proteins in Other Bacterial Pathogens. Clin. Microbiol. Rev. 18: 102-127 [Abstract] [Full Text]  
  • Turner, M. S., Hafner, L. M., Walsh, T., Giffard, P. M. (2004). Identification and Characterization of the Novel LysM Domain-Containing Surface Protein Sep from Lactobacillus fermentum BR11 and Its Use as a Peptide Fusion Partner in Lactobacillus and Lactococcus. Appl. Environ. Microbiol. 70: 3673-3680 [Abstract] [Full Text]  


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