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

Geldart, Kathryn; Forkus, Brittany; McChesney, Evelyn; McCue, Madeline; Kaznessis, Yiannis N.

doi:10.3390/ph9040060

Cited by 25 publications

(22 citation statements)

References 51 publications

(64 reference statements)

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“…We thus transformed pMPES2 and pMPES2:A, B, H, and BHA into EcN to generate respectively a control strain, a strain producing Enterocin A, Enterocin B, and Hiracin JM79 individually, and a strain producing all three peptides. Enterocin A and Hiracin JM79 were selected because they were the most potent bacteriocins targeting VRE in the original pMPES system . Enterocin B was selected because of previous evidence that it may act synergistically or via a different mechanism of action with Enterocin A …”

Section: Resultsmentioning

confidence: 99%

“…34,35 It was previously found that while the original pMPES was functional, its activity could be drastically improved by minor changes in the RBS. 29 It has been shown that the translation efficiency is significantly impacted by the genetic sequence immediately up and downstream of the RBS. 36,37 For the upstream sequence, we used the sequence produced by the the Salis Laboratory's RBS Calculator which is generated as part of the RBS optimization.…”

Section: Plasmid Design and Constructionmentioning

confidence: 99%

“…Enterocin A and Hiracin JM79 were selected because they were the most potent bacteriocins targeting VRE in the original pMPES system. 29 Enterocin B was selected because of previous evidence that it may act synergistically or via a different mechanism of action with Enterocin A. 38 As mentioned above, all peptides were expressed as a fusion of the Vsp tag and the mature bacteriocin.…”

Section: Plasmid Design and Constructionmentioning

confidence: 99%

“…We previously developed a peptide expression vector for EcN that employs the Microcin V secretion system to secrete a variety of bacteriocins of Gram‐positive and Gram‐negative origin . We showed that this modular peptide secretion system (pMPES) could secrete inhibitory concentrations of four bacteriocins with activity against VRE.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Engineered E. coli Nissle 1917 for the reduction of vancomycin‐resistant Enterococcus in the intestinal tract

Geldart

Kommineni

Forbes

et al. 2018

Bioengineering & Transla Med

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Vancomycin‐resistant Enterococcus (VRE) poses a serious threat in hospitals where they densely colonize the intestinal tracts of patients. In vulnerable hosts, these pathogens may translocate to the bloodstream and become lethal. The ability to selectively reduce VRE in the intestinal tracts of patients could potentially prevent many of these translocation events and reduce the spread of the pathogen. Herein, we have engineered Escherichia. coli Nissle 1917 to produce and secrete three antimicrobial peptides, Enterocin A, Enterocin B, and Hiracin JM79, to specifically target and kill Enterococcus. These peptides exhibited potent activity against both Enterococcus faecium and Enterococcus faecalis, the two most prominent species responsible for VRE infections. We first discuss the optimization of the system used to express and secrete the peptides. We then show that by simultaneously expressing these peptides, both E. faecium and E. faecalis were drastically inhibited. We then demonstrate a suppression of the development of resistance when supernatant from the E. coli producer strains was used to treat E. faecium. Finally, we tested the efficacy of the probiotic in a VRE colonization model in mice. These studies showed that administration of the engineered probiotic significantly reduced the levels of both E. faecium and E. faecalis in the feces of male Balb/cJ mice.

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Section: Resultsmentioning

confidence: 99%

Section: Plasmid Design and Constructionmentioning

confidence: 99%

Section: Plasmid Design and Constructionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Engineered E. coli Nissle 1917 for the reduction of vancomycin‐resistant Enterococcus in the intestinal tract

Geldart

Kommineni

Forbes

et al. 2018

Bioengineering & Transla Med

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“…Prolonged release of AMPs by engineered probiotics may kill commensal bacteria, and thus mitigate their beneficial effects. Nevertheless, as many as seven AMPs have been expressed in a recently reported modular peptide expression system pMPES [77]. In addition, commensal Enterococcus faecalis strain has been engineered to express bacteriocin-21 (a variant enterocin AS-48) via a plasmid with impaired bacterial conjugation ability to de-colonize vancomycin-resistant enterococci in the mouse gut [78**,79].…”

Section: Application Strategies Of Antimicrobial Peptidesmentioning

confidence: 99%

Host defense antimicrobial peptides as antibiotics: design and application strategies

Mishra

Reiling

Zarena

et al. 2017

Current Opinion in Chemical Biology

246

209

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This review deals with the design and application strategies of new antibiotics based on naturally occurring antimicrobial peptides (AMPs). The initial candidate can be designed based on three-dimensional structure or selected from a library of peptides from natural or laboratory sources followed by optimization via structure-activity relationship studies. There are also advanced application strategies such as induction of AMP expression from host cells by various factors (e.g., metals, amino acids, vitamin D and sunlight), the use of engineered probiotic bacteria to deliver peptides, the design of prodrug and peptide conjugates to improve specific targeting. In addition, combined uses of newly developed AMPs with existing antimicrobial agents may provide a practical avenue for effective management of antibiotic-resistant bacteria (superbugs, including biofilm). Finally, we highlight AMPs already in use or under clinical trials.

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Multispecies activity screening of microcin J25 mutants yields antimicrobials with increased specificity toward pathogenic Salmonella species relative to human commensal Escherichia coli

Ritter

Yang

Kaznessis

et al. 2018

Biotech & Bioengineering

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Modern large-scale agricultural practices that incorporate high density farming with subtherapeutic antibiotic dosing are considered a major contributor to the rise of antibiotic-resistant bacterial infections of humans with species of Salmonella being a leading agriculture-based bacterial infection. Microcin J25, a potent and highly stable antimicrobial peptide active against Enterobacteriaceae, is a candidate antimicrobial against multiple Salmonella species. Emerging evidence supports the hypothesis that the composition of the microbiota of the gastrointestinal tract prevents a variety of diseases by preventing infectious agents from proliferating. Reducing clearance of off-target bacteria may decrease susceptibility to secondary infection. Of the Enterobacteriaceae susceptible to microcin J25, Escherichia coli are the most abundant within the human gut. To explore the modulation of specificity, a collection of 207 mutants encompassing 12 positions in both the ring and loop of microcin J25 was built and tested for activity against Salmonella and E. coli strains. As has been found previously, mutational tolerance of ring residues was lower than loop residues, with 22% and 51% of mutations, respectively, retaining activity toward at least one target within the target organism test panel. The multitarget screening elucidated increased mutational tolerance at position G2, G3, and G14 than previously identified in panels composed of single targets. Multiple mutations conferred differential response between the different targets. Examination of specificity differences between mutants found that 30% showed significant improvements to specificity toward any of the targets. Generation and testing of a combinatorial library designed from the point-mutant study revealed that microcin J25 reduces off-target activity toward commensal human-derived E. coli isolates by 81% relative to Salmonella enterica serovar Enteritidis. These in vitro specificity improvements are likely to improve in vivo treatment efficacy by reducing clearance of commensal bacteria in the gastrointestinal tract of hosts.

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

Cited by 25 publications

References 51 publications

Engineered E. coli Nissle 1917 for the reduction of vancomycin‐resistant Enterococcus in the intestinal tract

Engineered E. coli Nissle 1917 for the reduction of vancomycin‐resistant Enterococcus in the intestinal tract

Host defense antimicrobial peptides as antibiotics: design and application strategies

Multispecies activity screening of microcin J25 mutants yields antimicrobials with increased specificity toward pathogenic Salmonella species relative to human commensal Escherichia coli

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