Production of active pediocin PA-1 in Escherichia coli using a thioredoxin gene fusion expression approach: cloning, expression, purification, and characterization.
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.
Beaulieu
Lucie
L
Biotechnology Research Institute, National Research Council, 6100 Royalmount Avenue, Montréal, QC 4P 2R2, Canada.
Tolkatchev
Dmitri
D
Jetté
Jean-François
JF
Groleau
Denis
D
Subirade
Muriel
M
D001430:Bacteriocins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D050939:Gene Fusion; D000070017:Pediocins; D010376:Pediococcus / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism; D013879:Thioredoxins / Q000235:genetics* / Q000378:metabolism
D001430:Bacteriocins; D000070017:Pediocins; D011994:Recombinant Proteins; C074288:pediocin PA-1; D013879:Thioredoxins