Improved production of class I lanthipeptides in <i>Escherichia coli</i>

Lee, Hyunji; Wu, Chunyu; Desormeaux, Emily; Sarksian, Raymond; Donk, Wilfred A. van der

doi:10.1039/d2sc06597e

Cited by 14 publications

(20 citation statements)

References 62 publications

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“…S2A). 16 In the current version, this platform includes a pETDuet plasmid that encodes a His-tagged blauticin leader and core sequence. The blauticin leader sequence is recognized by blauticin-specific LanB and LanC ( bpcB and bpcC gene, respectively) encoded in the pCDFDuet plasmid.…”

Section: Resultsmentioning

confidence: 99%

“…14,15 Using this information, an improved production platform has been established for facile expression and production of class I lanthipeptides. 16 Blauticin, a class I lantibiotic produced by the gut commensal bacteria Blautia producta SCSK (BPSCSK), exerts colonization resistance and clearance of Vancomycin-resistant Enterococci (VRE) in vivo. 17 A previous study showed that the in vivo VRE colonization in colon was inhibited by BPSCSK but not by Lactococcus lactis, the producing strain of the well-studied class I lantibiotic nisin.…”

Section: Introductionmentioning

confidence: 99%

“…14, 15 Using this information, an improved production platform has been established for facile expression and production of class I lanthipeptides. 16…”

Section: Introductionmentioning

confidence: 99%

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Structure-Activity Relationship Studies of Novel Gut-derived Lantibiotics Against Human Gut Commensals

Zhang,

Wu,

Dorantes

et al. 2023

Preprint

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Recent advances in sequencing techniques unveiled the vast potential of ribosomally synthesized and post-translationally modified peptides (RiPPs) encoded in microbiomes. Class I lantibiotics such as nisin A, widely used as a food preservative, have been investigated for their efficacy in killing pathogens. However, the impact of nisin and nisin-like class I lantibiotics on commensal bacteria residing in the human gut remains unclear. Here, we report six gut-derived class I lantibiotics that are close homologs of nisin, four of which are novel. We applied an improved lantibiotic expression platform to produce and purify these lantibiotics for antimicrobial assays. We determined their minimal inhibitory concentration (MIC) against both Gram-positive human pathogens and gut commensals, and profiled the lantibiotic resistance genes in these pathogens and commensals. SAR studies with variants revealed key regions and residues that impact their antimicrobial properties. Our characterization and SAR studies of nisin-like lantibiotics against both pathogens and human gut commensals could shed light on the future development of lantibiotic-based therapeutics and food preservatives.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Structure-Activity Relationship Studies of Novel Gut-derived Lantibiotics Against Human Gut Commensals

Zhang,

Wu,

Dorantes

et al. 2023

Preprint

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“…Glutathione (GSH) adducts to dehydroamino acids (Dha/Dhb) have been observed previously during heterologous production of lanthipeptides in E. coli. [59][60][61][62] LanC-like proteins (LanCL) have been shown to add GSH to peptides in mammalian cells, and indeed in vitro LanCL also adds GSH to CylLL. 63 To determine the location of the GSH adduct produced in HEK293 cells, a chymotrypsin digest was performed followed by matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS).…”

Section: Expression Of Lanthipeptides In Mammalian Cellsmentioning

confidence: 99%

Expression of Lanthipeptides in Human Cells

Eslami,

Rahman,

van der Donk

2023

Preprint

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Cyclic peptides represent a burgeoning area of interest in therapeutic and biotechnological research. In opposition to their linear counterparts, cyclic peptides, such as certain ribosomally synthesized and post-translationally modified peptides (RiPPs), are more conformationally constrained and less susceptible to proteolytic degradation. The lanthipeptide RiPP cytolysin L forms a covalently enforced helical structure that may be used to disrupt helical interactions at protein-protein interfaces. Herein, an expression system is reported to produce lanthipeptides and structurally diverse cytolysin L derivatives in mammalian cells. Successful targeting of lanthipeptides to the nucleus is demonstrated. In vivo expression and targeting of such peptides in mammalian cells may allow for screening of lanthipeptide inhibitors of native protein-protein interactions.

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“…Previous studies have demonstrated that introducing GluRS and tRNA Glu from the native lanthipeptide-producing organism could improve the production of fully modified peptides in E. coli. , Using this information, an improved production platform has been established for facile production of class I lanthipeptides in E. coli …”

Section: Introductionmentioning

confidence: 99%

Activity of Gut-Derived Nisin-like Lantibiotics against Human Gut Pathogens and Commensals

Zhang,

Wu,

Moreira

et al. 2024

ACS Chem. Biol.

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Recent advances in sequencing techniques unveiled the vast potential of ribosomally synthesized and post-translationally modified peptides (RiPPs) encoded in microbiomes. Class I lantibiotics such as nisin A, widely used as a food preservative, have been investigated for their efficacy in killing pathogens. However, the impact of nisin and nisin-like class I lantibiotics on commensal bacteria residing in the human gut remains unclear. Here, we report six gut-derived class I lantibiotics that are close homologues of nisin, four of which are novel. We applied an improved lantibiotic expression platform to produce and purify these lantibiotics for antimicrobial assays. We determined their minimal inhibitory concentration (MIC) against both Gram-positive human pathogens and gut commensals and profiled the lantibiotic resistance genes in these pathogens and commensals. Structure−activity relationship (SAR) studies with analogs revealed key regions and residues that impact their antimicrobial properties. Our characterization and SAR studies of nisin-like lantibiotics against both pathogens and human gut commensals could shed light on the future development of lantibiotic-based therapeutics and food preservatives.

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Improved production of class I lanthipeptides in Escherichia coli

Abstract: Lanthipeptides are ribosomally synthesised and post-translationally modified peptides containing lanthionine (Lan) and methyllanthionine (MeLan) residues that are formed by dehydration of Ser/Thr residues followed by conjugate addition of Cys to...

Cited by 14 publications

References 62 publications

Structure-Activity Relationship Studies of Novel Gut-derived Lantibiotics Against Human Gut Commensals

Structure-Activity Relationship Studies of Novel Gut-derived Lantibiotics Against Human Gut Commensals

Expression of Lanthipeptides in Human Cells

Activity of Gut-Derived Nisin-like Lantibiotics against Human Gut Pathogens and Commensals

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