Recombinant pediocin in <scp><i>L</i></scp><i>actococcus lactis</i>: increased production by propeptide fusion and improved potency by co‐production with <scp>PedC</scp>

Back, Alexandre; Borges, Frédéric; Mangavel, Cécile; Paris, Cédric; Rondags, Emmanuel; Kapel, Romain; Aymes, Arnaud; Rogniaux, Hélène; Pavlović, Marija; Heel, Auke J. van; Kuipers, Oscar P.; Revol-Junelles, Anne-Marie; Cailliez‐Grimal, Catherine

doi:10.1111/1751-7915.12285

Cited by 11 publications

(5 citation statements)

References 71 publications

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“…The functional expression is noteworthy because of the presence of intramolecular disulfide bonds which are less common in proteins of Gram‐positive LAB (Davey et al ., ). Among recombinant proteins, disulfide bond‐containing bacteriocins have been produced in L. lactis , and evasins may share the same mechanisms of disulfide bond formation (Back et al ., ; Davey et al ., ).…”

Section: Discussionmentioning

confidence: 99%

Evasin‐displaying lactic acid bacteria bind different chemokines and neutralize CXCL8 production in Caco‐2 cells

Škrlec

Janež

Rogelj

et al. 2017

Microbial Biotechnology

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SummaryChemokines are key signals in the immune system and play an important role as proinflammatory mediators in the pathology of inflammatory bowel disease and colorectal cancer, making them an important target for therapy. Recombinant lactic acid bacteria (LAB) were engineered to bind CC and CXC chemokines by displaying chemokine‐binding proteins evasin‐1, evasin‐3 and evasin‐4 on their surface. Evasin genes were cloned into lactococcal surface display vector and overexpressed in L. lactis NZ9000 and NZ9000ΔhtrA in fusion with secretion signal and surface anchor. Evasin‐displaying bacteria removed from 15% to 90% of 11 different chemokines from the solution as determined with ELISA and Luminex multiplexing assays, whereby L. lactis NZ9000ΔhtrA proved more efficient. Lactobacillus salivarius ATCC 11741 was coated with L. . lactis‐expressed evasin fusion protein, and its ability to bind chemokines was also confirmed. Evasin‐3‐displaying L. lactis removed 76.0% of IL‐1β‐induced CXCL8 from the supernatant of Caco‐2 epithelial cells. It also prevented secretion of CXCL8 from Caco‐2 cells in a time‐dependent manner when added before induction with IL‐1β. Evasin‐displaying LAB have the ability to bind multiple chemokines simultaneously and exert synergistic activity. This innovative treatment approach therefore has the potential for mucosal therapy of inflammatory bowel disease or colorectal cancer.

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Section: Discussionmentioning

confidence: 99%

Evasin‐displaying lactic acid bacteria bind different chemokines and neutralize CXCL8 production in Caco‐2 cells

Škrlec

Janež

Rogelj

et al. 2017

Microbial Biotechnology

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“…This strategy is essential for bacteriocins containing post-translational modifications such as nisin but might not be required for linear and non-modified peptides. In fact, it was shown for several bacteriocins that the double glycine SPs can be successfully replaced by Sec-dependent SPs ( McCormick et al, 1996 ; Martín et al, 2007 ; Borrero et al, 2011 ; Back et al, 2016 ) and recently a fusion of the SP of lactococcal protein Usp45 and GarQ allowed recombinant GarQ production by L. lactis ( Feito et al, 2023 ). To identify a suitable Sec-dependent SP for GarQ secretion by C. glutamicum we screened an available library of B. subtilis SPs.…”

Section: Discussionmentioning

confidence: 99%

Optimized recombinant production of the bacteriocin garvicin Q by Corynebacterium glutamicum

Desiderato,

Müller,

Schretzmeier

et al. 2024

Front. Microbiol.

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Bacteriocins are antimicrobial peptides applied in food preservation and are interesting candidates as alternatives to conventional antibiotics or as microbiome modulators. Recently, we established Corynebacterium glutamicum as a suitable production host for various bacteriocins including garvicin Q (GarQ). Here, we establish secretion of GarQ by C. glutamicum via the Sec translocon achieving GarQ titers of about 7 mg L–1 in initial fermentations. At neutral pH, the cationic peptide is efficiently adsorbed to the negatively charged envelope of producer bacteria limiting availability of the bacteriocin in culture supernatants. A combination of CaCl2 and Tween 80 efficiently reduces GarQ adsorption to C. glutamicum. Moreover, cultivation in minimal medium supplemented with CaCl2 and Tween 80 improves GarQ production by C. glutamicum to about 15 mg L–1 but Tween 80 resulted in reduced GarQ activity at later timepoints. Using a reporter strain and proteomic analyses, we identified HtrA, a protease associated with secretion stress, as another potential factor limiting GarQ production. Transferring production to HtrA-deficient C. glutamicum K9 improves GarQ titers to close to 40 mg L–1. Applying conditions of low aeration prevented loss in activity at later timepoints and improved GarQ titers to about 100 mg L–1. This is about 50-fold higher than previously shown with a C. glutamicum strain employing the native GarQ transporter GarCD for secretion and in the range of levels observed with the native producer Lactococcus petauri B1726. Additionally, we tested several synthetic variants of GarQ and were able to show that exchange of the methionine in position 5 to a phenylalanine (GarQM5F) results in markedly increased activity against Lactococcus lactis and Listeria monocytogenes. In summary, our findings shed light on several aspects of recombinant GarQ production that may also be of relevance for production with natural producers and other bacteriocins.

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“…Annotation performed on the MicroScope platform ( 8 ) detected 2,589 coding sequences (CDSs) and 38 tRNAs. Upstream from the dvnVTIRK gene cluster, which was previously reported as being responsible for divercin V41 production ( 4 ), we noticed a CDS similar to PedC, a thiol-disulfide oxidoreductase putatively involved in the posttranslational maturation of bacteriocins ( 9 ). No other bacteriocin production genes were identified.…”

Section: Genome Announcementmentioning

confidence: 76%

Draft Genome Sequence of Carnobacterium divergens V41, a Bacteriocin-Producing Strain

et al. 2016

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Recombinant pediocin in Lactococcus lactis: increased production by propeptide fusion and improved potency by co‐production with PedC

Cited by 11 publications

References 71 publications

Evasin‐displaying lactic acid bacteria bind different chemokines and neutralize CXCL8 production in Caco‐2 cells

Evasin‐displaying lactic acid bacteria bind different chemokines and neutralize CXCL8 production in Caco‐2 cells

Optimized recombinant production of the bacteriocin garvicin Q by Corynebacterium glutamicum

Draft Genome Sequence of Carnobacterium divergens V41, a Bacteriocin-Producing Strain

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