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Applied and Environmental Microbiology, August 2005, p. 4263-4268, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4263-4268.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Simultaneous Display of Multiple Foreign Peptides in the FliD Capping and FliC Filament Proteins of the Escherichia coli Flagellum

Katariina Majander, Timo K. Korhonen, and Benita Westerlund-Wikström*

General Microbiology, Faculty of Biosciences, University of Helsinki, Helsinki, Finland

Received 16 December 2004/ Accepted 3 March 2005

The bacterial flagellum is composed of more than 20 different proteins. The filament, which constitutes the major extracellular part of the flagellum, is built up of approximately 20,000 FliC molecules that assemble at the growing distal end of the filament. A capping structure composed of five FliD molecules located at the tip of the filament promotes polymerization of FliC. Lack of FliD leads to release of the subunits into the growth medium. We show here that FliD can be successfully used in bacterial surface display. We tested various insertion sites in the capping protein, and the optimal region for display was at the variable region in FliD. Deletion and/or insertion at other sites resulted in decreased formation of flagella. We further developed the technique into a multihybrid display system in which three foreign peptides are simultaneously expressed within the same flagellum, i.e., D repeats of FnBPA from Staphylococcus aureus at the tip and fragments of YadA from Yersinia enterocolitica as well as SlpA from Lactobacillus crispatus along the filament. This technology can have biotechnological applications, e.g., in simultaneous delivery of several effector molecules.

* Corresponding author. Mailing address: General Microbiology, Faculty of Biosciences, P.O. Box 56 (Viikinkaari 9C), FIN-00014 University of Helsinki, Finland. Phone: 358-9-19159251. Fax: 358-9-191 59262. E-mail: benita.westerlund{at}helsinki.fi.

Applied and Environmental Microbiology, August 2005, p. 4263-4268, Vol. 71, No. 8
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.8.4263-4268.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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