alexa Bacillus subtilis Spore Surface Display System Protects
ISSN: 2155-952X

Journal of Biotechnology & Biomaterials
Open Access

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Research Article

Bacillus subtilis Spore Surface Display System Protects Recombinant Proteins from Degradation-Verified Hypothesis

Tomasz Łęga1*, Paulina Weiher2, Monika Paszkiewicz3 and Dawid Nidzworski2,4

1Department of Medical Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk, Dębinki 1, 80-211 Gdańsk, Poland

2Department of Recombinant Vaccine, Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk, Kładki 24, 80-822 Gdańsk, Poland

3Department of Environmental Analytics, Faculty of Chemistry, University of Gdansk, ul. Wita Stwosza 63, 80-308 Gdansk, Poland

4Institute of Biotechnology and Molecular Medicine, Trzy Lipy 3 St., 80-172 Gdańsk

*Corresponding Author:
Łęga T
Department of Medical Biotechnology
Intercollegiate Faculty of Biotechnology
University of Gdańsk and Medical University of Gdańsk
Dębinki 1, 80-211 Gdańsk, Poland
Tel: +48725108632
E-mail: [email protected]

Received date: May 19, 2017; Accepted date: June 24, 2017; Published date: June 30, 2017

Citation: Łęga T, Weiher P, Paszkiewicz M, Nidzworski D (2017) Bacillus subtilis Spore Surface Display System Protects Recombinant Proteins from Degradation- Verified Hypothesis. J Biotechnol Biomater 7: 263. doi: 10.4172/2155-952X.1000263

Copyright: © 2017 Łęga T, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

 

Abstract

Endospores of Bacillus subtilis have been used extensively as a platform for recombinant protein display for nearly two decades. Main use of the spore surface display system is generation of oral vaccines against many human and animal pathogens. Formulation of oral vaccine based on spores is an attractive approach to alternative vaccination due to the timesaving and relative easiness of production. Another advantage of such formulation is stability of presented antigens. It is assumed that the spore coat structure prevents degradation of displayed proteins by many destructive factors such as heat, proteases or stomach environment. However, there is little scientific background and substantial lack of experiments verifying this statement. In our work, we tested protective properties of spores against degradation of displayed antigens in harsh environment. We constructed B. subtilis strains producing spores presenting highly conserved long α-helix (LAH) region of the influenza A virus hemagglutinin. The constructs were obtained by fusion of LAH antigen to protein CotB or CotZ from the spore coat. We treated recombinant spores with destructive agents such as heat, protease, low pH and high-energy irradiation to test protective features of the spore coat. After treatment, spore coat extracts were analyzed by western blot to study fate of the displayed antigen. Results that we publish indicate that spore coat protects displayed antigen from degradation. This work is a strong support of hypothesis stating protective properties of spore surface display system against antigen degradation.

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