Production of active pediocin PA-1 in Escherichia coli using a thioredoxin gene fusion expression approach: cloning, expression, purification, and characterization

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1139/W07-089
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Name affiliation
  1. National Research Council Canada. NRC Biotechnology Research Institute
FormatText
TypeArticle
Journal titleCanadian Journal of Microbiology
Volume53
Issue11
Pages1246–1258; # of pages: 13
SubjectBacteria; bacteriocins; biotechnology; biotechnology; cells; escherichia coli; food preservatives; pediococcus; peptides; pharmaceutical; protein; pediocin PA-1; pedA expression; thioredoxin; biological activity; expression de la pedA; Escherichia coli; thiorédoxine; activité biologique
Abstract
Antimicrobial peptides possess cationic and amphipathic properties that allow for interactions with the membrane of living cells. Bacteriocins from lactic acid bacteria, in particular, are currently being studied for their potential use as food preservatives and for applications in health care. However, bacteriocin exploitation is often limited owing to low production yields. Gene cloning and heterologous protein or peptide production is one way to possibly achieve overexpression of bacteriocins to support biochemical studies. In this work, production of recombinant active pediocin PA-1 (PedA) was accomplished in Escherichia coli using a thioredoxin (trx) gene fusion (trx–pedA) expression approach. Trx–PedA itself did not show any biological activity, but upon cleavage by an enterokinase, biologically active pediocin PA-1 was obtained. Recombinant pediocin PA-1 characteristics (molecular mass, biological activity, physicochemical properties) were very similar to those of native pediocin PA-1. In addition, a 4- to 5-fold increase in production yield was obtained, by comparison with the PA-1 produced naturally by Pediococcus acidilactici PAC 1.0. The new production method, although not optimized, offers great potential for supporting further investigations on pediocin PA-1 and as a first-generation process for the production of pediocin PA-1 for high-value applications.
Publication date
LanguageEnglish
NRC number47806
47806
NPARC number3539960
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