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

Paper ID Volume ID Publish Year Pages File Format Full-Text
25475 43576 2007 12 PDF Available
Title
Heterologous production and secretion of Clostridium perfringens β-toxoid in closely related Gram-positive hosts
Abstract

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.

Keywords
Clostridium perfringens β-toxin; Bacillus subtilis; Lactococcus lactis; Heterologous protein expression; Protein secretion; rpsF promoter; Vaccine production
First Page Preview
Heterologous production and secretion of Clostridium perfringens β-toxoid in closely related Gram-positive hosts
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 127, Issue 3, 10 January 2007, Pages 361–372
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering