title: Heterologous production and secretion of Clostridium perfringens beta-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 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.
authors.affiliation: Department of Genetics, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, NL-9751 NN Haren, The Netherlands.
authors.forename: Reindert
authors.initials: R
authors.lastname: Nijland
authors.forename: Cordula
authors.initials: C
authors.lastname: Lindner
authors.forename: Mariska
authors.initials: M
authors.lastname: van Hartskamp
authors.forename: Leendert W
authors.initials: LW
authors.lastname: Hamoen
authors.forename: Oscar P
authors.initials: OP
authors.lastname: Kuipers
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D001428:Bacterial Vaccines / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D003016:Clostridium perfringens / Q000235:genetics* / Q000276:immunology; D015870:Gene Expression; D006094:Gram-Positive Bacteria / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D014121:Toxoids / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D001428:Bacterial Vaccines; C035454:Clostridium perfringens toxoid; D011994:Recombinant Proteins; D014121:Toxoids