title: Recombinant pediocin in Lactococcus lactis: increased production by propeptide fusion and improved potency by co-production with PedC.
abstract: We describe the impact of two propeptides and PedC on the production yield and the potency of recombinant pediocins produced in Lactococcus 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.
authors.affiliation: Laboratoire d'Ingénierie des Biomolécules (LIBio), ENSAIA, Université de Lorraine, 2 Avenue de la Forêt de Haye, Vandœuvre-lès-Nancy, 54518, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Back
authors.affiliation: Laboratoire d'Ingénierie des Biomolécules (LIBio), ENSAIA, Université de Lorraine, 2 Avenue de la Forêt de Haye, Vandœuvre-lès-Nancy, 54518, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Borges
authors.affiliation: Laboratoire d'Ingénierie des Biomolécules (LIBio), ENSAIA, Université de Lorraine, 2 Avenue de la Forêt de Haye, Vandœuvre-lès-Nancy, 54518, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Mangavel
authors.affiliation: Laboratoire d'Ingénierie des Biomolécules (LIBio), ENSAIA, Université de Lorraine, 2 Avenue de la Forêt de Haye, Vandœuvre-lès-Nancy, 54518, France.
authors.forename: Cédric
authors.initials: C
authors.lastname: Paris
authors.affiliation: Laboratoire Réactions et Génie des Procédés (LRGP), CNRS-UMR 7274, Université de Lorraine, 2 Avenue de la Forêt de Haye, Vandœuvre-lès-Nancy, 54518, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Rondags
authors.affiliation: Laboratoire Réactions et Génie des Procédés (LRGP), CNRS-UMR 7274, Université de Lorraine, 2 Avenue de la Forêt de Haye, Vandœuvre-lès-Nancy, 54518, France.
authors.forename: Romain
authors.initials: R
authors.lastname: Kapel
authors.affiliation: Laboratoire Réactions et Génie des Procédés (LRGP), CNRS-UMR 7274, Université de Lorraine, 2 Avenue de la Forêt de Haye, Vandœuvre-lès-Nancy, 54518, France.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Aymes
authors.affiliation: INRA Unité Biopolymères Interactions Assemblages (UR1268), Rue de la Géraudière, Nantes, 44316, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Rogniaux
authors.affiliation: INRA Unité Biopolymères Interactions Assemblages (UR1268), Rue de la Géraudière, Nantes, 44316, France.
authors.forename: Marija
authors.initials: M
authors.lastname: Pavlović
authors.affiliation: Department of Molecular Genetics, GBB Institute, University of Gronningen, Nijenborgh 7, 9747AG, Groningen, The Netherlands.
authors.forename: Auke J
authors.initials: AJ
authors.lastname: van Heel
authors.affiliation: Department of Molecular Genetics, GBB Institute, University of Gronningen, Nijenborgh 7, 9747AG, Groningen, The Netherlands.
authors.forename: Oscar P
authors.initials: OP
authors.lastname: Kuipers
authors.affiliation: Laboratoire d'Ingénierie des Biomolécules (LIBio), ENSAIA, Université de Lorraine, 2 Avenue de la Forêt de Haye, Vandœuvre-lès-Nancy, 54518, France.
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Revol-Junelles
authors.affiliation: Laboratoire d'Ingénierie des Biomolécules (LIBio), ENSAIA, Université de Lorraine, 2 Avenue de la Forêt de Haye, Vandœuvre-lès-Nancy, 54518, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Cailliez-Grimal
mesh_terms: D004797:Enzyme-Linked Immunosorbent Assay; D013294:Lactococcus lactis / Q000235:genetics* / Q000378:metabolism*; D060847:Metabolic Engineering; D053858:Metabolic Networks and Pathways / Q000235:genetics; D008826:Microbial Sensitivity Tests; D000070017:Pediocins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D017422:Sequence Analysis, DNA
chemical_list: D000070017:Pediocins; D011994:Recombinant Proteins