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

Reindert Nijland, Cordula Lindner, Mariska van Hartskamp, Leendert W. Hamoen, Oscar P. Kuipers*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review


The spore forming bacterium Clostridium perfringens is a widely occurring pathogen. Vaccines against C. perfringens type B and C are currently manufactured using β-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 β-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 μg ml-1 culture. However, the β-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 β-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.

Original languageEnglish
Pages (from-to)361-372
Number of pages12
JournalJournal of Biotechnology
Issue number3
Publication statusPublished - 10 Jan 2007
Externally publishedYes


  • Bacillus subtilis
  • Clostridium perfringens β-toxin
  • Heterologous protein expression
  • Lactococcus lactis
  • Protein secretion
  • rpsF promoter
  • Vaccine production


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