Recombinant pediocin in Lactococcus lactis: increased production by propeptide fusion and improved potency by co‐production with PedC. Issue 4 (3rd July 2015)

• Record Type: Journal Article
• Authors: Back, Alexandre; Borges, Frédéric; Mangavel, Cécile; Paris, Cédric; Rondags, Emmanuel; Kapel, Romain; Aymes, Arnaud; Rogniaux, Hélène; Pavlović, Marija; van Heel, Auke J.; Kuipers, Oscar P.; Revol‐Junelles, Anne‐Marie; Cailliez‐Grimal, Catherine
• Is Part Of: Microbial biotechnology. Volume 9:Issue 4(2016:Jul.)
• Page Start: 466
• Page End: 477
• Publication Date: 2015-07-03
• Subjects: Microbial biotechnology -- Periodicals; Biotechnology; Microbiology; 660.62
• DOI: 10.1111/1751-7915.12285
• Languages: English
• ISSNs: 1751-7915

Abstract

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 Usp45 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. Abstract : We show that heterologous production of pediocin in Lactococcus lactis can be improved both quantitatively and qualitatively. The amount of produced pediocin can be increased by inserting the propeptide SD or LEISSTCDA between the signal peptide and the structural mature bacteriocin. The potency of the bacteriocin can be improved by co‐expressing the putative Thiol‐Disulfide Oxidoreductase (TDOR) PedC.