Heterologous production and secretion of Clostridium perfringens beta-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

30 Citations (Scopus)
254 Downloads (Pure)

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 mu g 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. (c) 2006 Elsevier B.V. All rights reserved.

Original languageEnglish
Pages (from-to)361-372
Number of pages12
JournalJournal of Biotechnology
Volume127
Issue number3
DOIs
Publication statusPublished - 10-Jan-2007

Keywords

  • Clostridium perfringens beta-toxin
  • Bacillus subtilis
  • Lactococcus lactis
  • heterologous protein expression
  • protein secretion
  • rpsF promoter
  • vaccine production
  • CONTROLLED GENE-EXPRESSION
  • BACILLUS-SUBTILIS
  • LACTOCOCCUS-LACTIS
  • ALPHA-AMYLASE
  • SYSTEM NICE
  • PROTEIN
  • TRANSFORMATION
  • TOXIN
  • NISIN
  • STREPTOCOCCI

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