Open AccessRecombinant pediocin in L actococcus lactis : increased production by propeptide fusion and improved potency by co‐production with PedC
Author(s) -
Back Alexandre,
Borges Frédéric,
Mangavel Cécile,
Paris Cédric,
Rondags Emmanuel,
Kapel Romain,
Aymes Arnaud,
Rogniaux Hélène,
Pavlović Marija,
Heel Auke J.,
Kuipers Oscar P.,
RevolJunelles AnneMarie,
CailliezGrimal Catherine
Publication year - 2016
Publication title -
microbial biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.287
H-Index - 74
ISSN - 1751-7915
DOI - 10.1111/1751-7915.12285
Subject(s) - recombinant dna , potency , biochemistry , protein precursor , heterologous , signal peptide , microbiology and biotechnology , fusion protein , biology , heterologous expression , chemistry , gene , in vitro
Summary We describe the impact of two propeptides and PedC on the production yield and the potency of recombinant pediocins produced in L actococcus lactis . On the one hand, the sequences encoding the propeptides SD or LEISSTCDA were inserted between the sequence encoding the signal peptide of U sp45 and the structural gene of the mature pediocin PA ‐1. On the other hand, the putative thiol‐disulfide oxidoreductase PedC was coexpressed with pediocin. The concentration of recombinant pediocins produced in supernatants was determined by enzyme‐linked immunosorbent assay. The potency of recombinant pediocins was investigated by measuring the minimal inhibitory concentration by agar well diffusion assay. The results show that propeptides SD or LEISSTCDA lead to an improved secretion of recombinant pediocins with apparently no effect on the antibacterial potency and that PedC increases the potency of recombinant pediocin. To our knowledge, this study reveals for the first time that pediocin tolerates fusions at the N ‐terminal end. Furthermore, it reveals that only expressing the pediocin structural gene in a heterologous host is not sufficient to get an optimal potency and requires the accessory protein PedC . In addition, it can be speculated that PedC catalyses the correct formation of disulfide bonds in pediocin.