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Microb Cell Fact. 2016 Aug 11;15(1):137. doi: 10.1186/s12934-016-0532-5.

Heterologous expression of antigenic peptides in Bacillus subtilis biofilms.

Author information

Institute of Virology, University of Zurich, Winterthurerstrasse 266a, 8057, Zurich, Switzerland.
Institute of Anatomy, University of Zurich, Zurich, Switzerland.
rqmicro Ltd, ETH, Otto-Stern-Weg 7, 8093, Zurich, Switzerland.
Institute of Virology, University of Zurich, Winterthurerstrasse 266a, 8057, Zurich, Switzerland.



Numerous strategies have been developed for the display of heterologous proteins in the surface of live bacterial carriers, which can be used as vaccines, immune-modulators, cancer therapy or bioremediation. Bacterial biofilms have emerged as an interesting approach for the expression of proteins of interest. Bacillus subtilis is a well-described, endospore-forming organism that is able to form biofilms and also used as a probiotic, thus making it a suitable candidate for the display of heterologous proteins within the biofilm. Here, we describe the use of TasA, an important structural component of the biofilms formed by B. subtilis, as a genetic tool for the display of heterologous proteins.


We first engineered the fusion protein TasA-mCherry and showed that was widely deployed within the B. subtilis biofilms. A significant enhancement of the expression of TasA-mCherry within the biofilm was obtained when depleting both tasA and sinR genes. We subsequently engineered fusion proteins of TasA to antigenic peptides of the E. granulosus parasite, paramyosin and tropomyosin. Our results show that the antigens were well expressed within the biofilm as denoted by macrostructure complementation and by the detection of the fusion protein in both immunoblot and immunohistochemistry. In addition, we show that the recombinant endospores of B. subtilis preserve their biophysical and morphological properties.


In this work we provide strong evidence pointing that TasA is a suitable candidate for the display of heterologous peptides, such as antigens, cytokines, enzymes or antibodies, in the B. subtilis biofilms. Finally, our data portray that the recombinant endospores preserve their morphological and biophysical properties and could be an excellent tool to facilitate the transport and the administration.


Antigen; Bacillus subtilis; Biofilm; E. granulosus; Endospores; Heterologous protein; Paramyosin; TasA; Tropomyosin; mCherry

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