Purification and characterization of the antimicrobial peptide microcin N

Corsini, Gino; Karahanian, Eduardo; Tello, Mario; Fernández, Katia; Rivero, Daniel; Saavedra, José M.; Ferrer, Alonso

doi:10.1111/j.1574-6968.2010.02106.x

Cited by 24 publications

(28 citation statements)

References 27 publications

(42 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…MccV, MccL and McnN are natively produced by E. coli while the remaining peptides are produced by Enterococcus , a Gram-positive genus of bacteria [17,19,23,32,33]. Like MccV, the studied enterocins are considered class II bacteriocins, which generally lack major post-translational modifications and are commonly secreted by ABC-transporters using N-terminal secretion tags [34].…”

Section: Resultsmentioning

confidence: 99%

pMPES: A Modular Peptide Expression System for the Delivery of Antimicrobial Peptides to the Site of Gastrointestinal Infections Using Probiotics

Geldart

Forkus

McChesney³

et al. 2016

Pharmaceuticals

View full text Add to dashboard Cite

Antimicrobial peptides are a promising alternative to traditional antibiotics, but their utility is limited by high production costs and poor bioavailability profiles. Bacterial production and delivery of antimicrobial peptides (AMPs) directly at the site of infection may offer a path for effective therapeutic application. In this study, we have developed a vector that can be used for the production and secretion of seven antimicrobial peptides from both Escherichia coli MC1061 F’ and probiotic E. coli Nissle 1917. The vector pMPES (Modular Peptide Expression System) employs the Microcin V (MccV) secretion system and a powerful synthetic promoter to drive AMP production. Herein, we demonstrate the capacity of pMPES to produce inhibitory levels of MccV, Microcin L (MccL), Microcin N (McnN), Enterocin A (EntA), Enterocin P (EntP), Hiracin JM79 (HirJM79) and Enterocin B (EntB). To our knowledge, this is the first demonstration of such a broadly-applicable secretion system for AMP production. This type of modular expression system could expedite the development of sorely needed antimicrobial technologies.

show abstract

Section: Resultsmentioning

confidence: 99%

pMPES: A Modular Peptide Expression System for the Delivery of Antimicrobial Peptides to the Site of Gastrointestinal Infections Using Probiotics

Geldart

Forkus

McChesney³

et al. 2016

Pharmaceuticals

View full text Add to dashboard Cite

show abstract

“…Analysis of the NRPSs in gene cluster one revealed that this locus likely encodes proteins involved in the biosynthesis of a microcin (Table 2). SMA1574A shows 41 % identity with microcin N of E. coli 2424 (Corsini et al , 2010) and the NRPS proteins, SMA1572 and SMA1571, are likely involved in the biosynthesis of a modifying group added to the microcin peptide encoded by this gene cluster (Rebuffat, 2012). In contrast, gene clusters two and five appear to direct the synthesis of multiple siderophores (Table 2).…”

Section: Resultsmentioning

confidence: 99%

Role of the phosphopantetheinyltransferase enzyme, PswP, in the biosynthesis of antimicrobial secondary metabolites by Serratia marcescens Db10

2014

View full text Add to dashboard Cite

Phosphopantetheinyltransferase (PPTase) enzymes fulfil essential roles in primary and secondary metabolism in prokaryotes, archaea and eukaryotes. PPTase enzymes catalyse the essential modification of the carrier protein domain of fatty acid synthases, polyketide synthases (PKSs) and non-ribosomal peptide synthetases (NRPSs). In bacteria and fungi, NRPS and PKS enzymes are often responsible for the biosynthesis of secondary metabolites with clinically relevant properties; these secondary metabolites include a variety of antimicrobial peptides. We have previously shown that in the Gram-negative bacterium Serratia marcescens Db10, the PPTase enzyme PswP is essential for the biosynthesis of an NRPS-PKS dependent antibiotic called althiomycin. In this work we utilize bioinformatic analyses to classify PswP as belonging to the F/KES subfamily of Sfp type PPTases and to putatively identify additional NRPS substrates of PswP, in addition to the althiomycin NRPS-PKS, in Ser. marcescens Db10. We show that PswP is required for the production of three diffusible metabolites by this organism, each possessing antimicrobial activity against Staphylococcus aureus. Genetic analyses identify the three metabolites as althiomycin, serrawettin W2 and an as-yet-uncharacterized siderophore, which may be related to enterobactin. Our results highlight the use of an individual PPTase enzyme in multiple biosynthetic pathways, each contributing to the ability of Ser. marcescens to inhibit competitor bacteria by the production of antimicrobial secondary metabolites.

show abstract

“…There are four known class IIa microcins, microcin V (MccV, previously named colicin V) (54, 55), microcin N (MccN, previously named Mcc24) (56), microcin L (MccL) (57), and microcin PDI (MccPDI) (58, 59). The operons encoding these microcins contain four or five genes, including the microcin precursor, immunity and export genes.…”

Section: Discussionmentioning

confidence: 99%

“…The operons encoding these microcins contain four or five genes, including the microcin precursor, immunity and export genes. MccN also encodes a putative regulator, with a histone-like nucleoid domain (56). The microcin precursor genes possess leader sequences of approximately 15 amino acids, containing the signature sequence MRXI/LX(9)GG/A (X=any amino acid), and are typically cleaved by the ABC transporters during export (60).…”

Section: Discussionmentioning

confidence: 99%

A putative microcin amplifies Shiga toxin 2a production ofEscherichia coliO157:H7

Figler¹,

Xiaoli

Banerjee³

et al. 2019

Preprint

View full text Add to dashboard Cite

word count: 315 15 Text word count: 5028 16 17 Abstract 24Escherichia coli O157:H7 is a foodborne pathogen, implicated in various multi-state 25 outbreaks. It encodes Shiga toxin on a prophage, and Shiga toxin production is linked to phage 26induction. An E. coli strain, designated 0.1229, was identified that amplified Stx2a production 27 when co-cultured with E. coli O157:H7 strain PA2. Growth of PA2 in 0.1229 cell-free 28 supernatants had a similar effect, even when supernatants were heated to 100°C for 10 min, but 29 not after treatment with Proteinase K. The secreted molecule was shown to use TolC for export 30 and the TonB system for import. The genes sufficient for production of this molecule were 31 localized to a 5.2 kb region of a 12.8 kb plasmid. This region was annotated, identifying 32 hypothetical proteins, a predicted ABC transporter, and a cupin superfamily protein. These genes 33were identified and shown to be functional in two other E. coli strains, and bioinformatic 34 analyses identified related gene clusters in similar and distinct bacterial species. These data 35 collectively suggest E. coli 0.1229 and other E. coli produce a microcin that induces the SOS 36 response in target bacteria. Besides adding to the limited number of microcins known to be 37 produced by E. coli, this study provides an additional mechanism by which stx2a expression is 38 increased in response to the gut microflora. 39 40

show abstract

Purification and characterization of the antimicrobial peptide microcin N

Cited by 24 publications

References 27 publications

pMPES: A Modular Peptide Expression System for the Delivery of Antimicrobial Peptides to the Site of Gastrointestinal Infections Using Probiotics

pMPES: A Modular Peptide Expression System for the Delivery of Antimicrobial Peptides to the Site of Gastrointestinal Infections Using Probiotics

Role of the phosphopantetheinyltransferase enzyme, PswP, in the biosynthesis of antimicrobial secondary metabolites by Serratia marcescens Db10

A putative microcin amplifies Shiga toxin 2a production ofEscherichia coliO157:H7

Contact Info

Product

Resources

About