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Heterologous production and secretion of Clostridium perfringens beta-toxoid in closely related Gram-positive hosts

Lookup NU author(s): Dr Reindert Nijland, Dr Leendert Hamoen

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Abstract

The spore forming bacterium Clostridium perfringens is a widely occurring pathogen. Vaccines against C. perfringens type B and C are currently manufactured using beta-toxin secreted by virulent C. perfringens strains. Large-scale production of vaccines from virulent strains requires stringent safety conditions and costly detoxification and control steps. Therefore, it would be beneficial to produce this toxin in a safe production host and in an immunogenic, but non-toxic form (toxoid). For high-level expression of beta-toxoid, we cloned the highly active ribosomal rpsF promoter of Bacillus subtilis in a broad host range multicopy plasmid. In B. subtilis, we obtained high intracellular production, up to 200 microg ml(-1) culture. However, the beta-toxoid was poorly secreted. The employed rpsF expression system allowed using the same expression plasmids in other heterologous hosts such as Lactococcus lactis and Streptococcus pneumoniae. In these organisms secretion of beta-toxoid was ten times higher compared to the best producing B. subtilis strain. These results show the usefulness of the rpsF based broad host range expression system.


Publication metadata

Author(s): Nijland R, Lindner C, van Hartskamp M, Hamoen LW, Kuipers OP

Publication type: Article

Journal: Journal of Biotechnology

Year: 2007

Volume: 127

Issue: 3

Pages: 361-72

ISSN (print): 0168-1656

ISSN (electronic): 1873-4863

Publisher: Elsevier BV

URL: http://dx.doi.org/10.1016/j.jbiotec.2006.07.014

DOI: 10.1016/j.jbiotec.2006.07.014

Notes: Journal Article Research Support, Non-U.S. Government Netherlands


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