Control of antimicrobial peptide synthesis by the agr quorum sensing system in Staphylococcus epidermidis: activity of the lantibiotic epidermin is regulated at the level of precursor peptide processing

Kies, Stefanie; Vuong, Cuong; Hille, Matthias; Peschel, Andreas; Meyer, Cornelia; Götz, Friedrich; Otto, Michael

doi:10.1016/s0196-9781(03)00046-9

Cited by 31 publications

(24 citation statements)

References 27 publications

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“…The two variants were present in different proportions in supernatants, with the larger one reaching only 10% of the spectrum intensity. These results were in agreement with those of a previous study reporting that two preepidermin fragments were detected in supernatants of an S. epidermidis ⌬epiP mutant strain (29). Furthermore, the strain producing truncated pregallidermins did not show self-toxicity even at higher concentrations than the ones expected in overexpression experiments (Fig.…”

Section: Discussionsupporting

confidence: 93%

“…Bacterial strains and plasmids are listed in Table 1. Escherichia coli strains were routinely grown in Luria-Bertani broth (46), whereas staphylococci were grown in B broth (29). All percentages refer to weight/volume unless otherwise specified.…”

Section: Methodsmentioning

confidence: 99%

“…The MS analysis was adapted from the method of Kies et al (29) and performed on a Finningan LCQ Deca LC/ESI-MS in cationic mode. A Bischoff Prontosil Eurobond C 18 5.0-m column was used for the stationary phase.…”

Section: Methodsmentioning

confidence: 99%

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Circumventing the Effect of Product Toxicity: Development of a Novel Two-Stage Production Process for the Lantibiotic Gallidermin

Valsesia

Medaglia

Held

et al. 2007

Appl Environ Microbiol

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Lantibiotics such as gallidermin are lanthionine-containing polypeptide antibiotics produced by grampositive bacteria that might become relevant for the treatment of various infectious diseases. So far, selftoxicity has prevented the isolation of efficient overproducing strains, thus hampering their thorough investigation and preventing their exploitation in fields other than the food area. We wanted to investigate the effect of lantibiotic precursor peptides on the producing strains in order to evaluate novel strategies for the overproduction of these promising peptides. In this study, gallidermin was chosen as a representative example of the type A lantibiotics. A Staphylococcus gallinarum Tü3928 mutant, whose gene for the extracellular pregallidermin protease GdmP was replaced by a kanamycin-resistance gene, was constructed. Mass spectrometry (MS) analysis indicated that this mutant produced fully posttranslationally modified gallidermin precursors with truncated versions of the leader peptide, but not the entire leader as predicted from the gdmA sequence. In filter-on-plate assays, these truncated pregallidermins showed no toxicity against Staphylococcus gallinarum Tü3928 up to a concentration of 8 g/liter (corresponding to approximately 2.35 mM), while gallidermin produced clear inhibitory zones at concentrations as low as 0.25 g/liter (0.12 mM). We showed that the lack of toxicity is due entirely to the presence of the truncated leader, since MS as well as bioassay analysis showed that the peptides resulting from tryptic cleavage of pregallidermins and gallidermin produced by S. gallinarum Tü3928 had identical masses and approximately the same specific activity. This demonstrates that even a shortened leader sequence is sufficient to prevent the toxicity of mature gallidermin. In nonoptimized fermentations, the gdmP mutant produced pregallidermin to a 50%-higher molar titer, suggesting that the absence of self-toxicity has a beneficial effect on gallidermin production and giving a first confirmation of the suitability of the overproduction strategy.Antibiotic resistance has spread dramatically among pathogens throughout the last two decades. Of particular concern is the increasing emergence of multiresistant pathogens in health care settings. Nowadays, about 70% of nosocomial infections in the United States are resistant to at least one antibiotic (35), and in 2002, the first clinical isolate of Staphylococcus aureus with high-level resistance to vancomycin, a drug of last resort, was isolated (8). As the number of resistant pathogens continues to grow, the number of new antibiotics to fight them is declining steeply (37). Therefore, the development of new antimicrobial agents should be encouraged.Type A lantibiotics are cationic, amphiphilic peptides that are produced by a wide range of gram-positive bacteria and show bactericidal activity against other gram-positive bacteria. They show a dual mode of action at nanomolar concentrations which involves pore formation and inhibition of peptidoglycan biosynth...

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Section: Discussionsupporting

confidence: 93%

Section: Methodsmentioning

confidence: 99%

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Circumventing the Effect of Product Toxicity: Development of a Novel Two-Stage Production Process for the Lantibiotic Gallidermin

Valsesia

Medaglia

Held

et al. 2007

Appl Environ Microbiol

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“…chromogenes is favoured during biofilm formation on the teat apex skin, whereby both biofilm formation and bacteriocin production are quorum sensing dependent (Kies et al . ; McDougald et al . ).…”

Section: Discussionmentioning

confidence: 99%

Antibacterial activities of coagulase-negative staphylococci from bovine teat apex skin and their inhibitory effect on mastitis-related pathogens

et al. 2014

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Aims: To explore antibacterial activities of coagulase-negative staphylococci (CoNS) from teat apices of dairy cows towards mastitis-causing pathogens. Methods and Results: Of 254 CoNS, 38 displayed bacteriocin-like activity after a first screening. Seven of these strains displayed activity against at least one mastitis-related pathogen (Streptococcus uberis, Streptococcus dysgalactiae and Staphylococcus aureus). Staphylococcus chromogenes L217 displayed the strongest inhibitory effect, being active against all tested mastitis-related pathogens and most tested CoNS. Based on cation exchange and reversed-phase chromatography, in addition to N-terminal Edman degradation and PCR, the antibacterial peptide was identified as a nukacin-type bacteriocin and named nukacin L217. Although staphylococcal bacteriocins are generally found in the cell-free supernatants of liquid cultures, Staph. chromogenes L217 only led to detectable activity when grown on agar medium. Conclusions: Bacteriocin-like activities are not uncommon among CoNS from teat apices and may inhibit mastitis-causing pathogens, as found for nukacin L217 production by Staph. chromogenes L217. Significance and Impact of the Study: Nukacin L217 is the first identified bacteriocin of the species Staph. chromogenes and displays unusual production kinetics, that is, requiring surface growth of its producer. The fact that nukacins are produced by different CoNS species suggests a role in the teat skin ecosystem.

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“…This may reflect the wide range of input signals that induce lantibiotic biosynthesis in the different producing organisms. Many lantibiotics regulate their own production in a cell density-dependent manner, frequently mediated by a two-component regulatory system (2), but this is often integrated with other external signals, such as developmental cues, pH, and cell stress responses (9,14,16,28). Deletion of mibA, encoding the microbisporicin prepropeptide, from Microbispora corallina NRRL 30420 markedly reduced mib gene expression, including that of the putative regulatory genes mibX and mibR, suggesting that microbisporicin similarly regulates its own biosynthesis (4).…”

Section: C-terminal S-[(z)-2-aminovinyl]-d-cysteinementioning

confidence: 99%

Feed-Forward Regulation of Microbisporicin Biosynthesis in Microbispora corallina

Foulston

Bibb

2011

J Bacteriol

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Lantibiotics are ribosomally synthesized, posttranslationally modified peptide antibiotics. Microbisporicin is a potent lantibiotic produced by the actinomycete Microbispora corallina and contains unique chlorinated tryptophan and dihydroxyproline residues. The biosynthetic gene cluster for microbisporicin encodes several putative regulatory proteins, including, uniquely, an extracytoplasmic function (ECF) factor, MibX , a likely cognate anti-factor, MibW, and a potential helix-turn-helix DNA binding protein, MibR. Here we examine the roles of these proteins in regulating microbisporicin biosynthesis. S1 nuclease protection assays were used to determine transcriptional start sites in the microbisporicin gene cluster and confirmed the presence of the likely ECF sigma factor ؊10 and ؊35 sequences in five out of six promoters. In contrast, the promoter of mibA, encoding the microbisporicin prepropeptide, has a typical Streptomyces vegetative sigma factor consensus sequence. The ECF sigma factor MibX was shown to interact with the putative anti-sigma factor MibW in Escherichia coli using bacterial two-hybrid analysis.MibX autoregulates its own expression but does not directly regulate expression of mibA. On the basis of quantitative reverse transcriptase PCR (qRT-PCR) data, we propose a model for the biosynthesis of microbisporicin in which MibR functions as an essential master regulator and the ECF sigma factor/anti-sigma factor pair, MibX /MibW, induces feed-forward biosynthesis of microbisporicin and producer immunity.

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Control of antimicrobial peptide synthesis by the agr quorum sensing system in Staphylococcus epidermidis: activity of the lantibiotic epidermin is regulated at the level of precursor peptide processing

Cited by 31 publications

References 27 publications

Circumventing the Effect of Product Toxicity: Development of a Novel Two-Stage Production Process for the Lantibiotic Gallidermin

Circumventing the Effect of Product Toxicity: Development of a Novel Two-Stage Production Process for the Lantibiotic Gallidermin

Antibacterial activities of coagulase-negative staphylococci from bovine teat apex skin and their inhibitory effect on mastitis-related pathogens

Feed-Forward Regulation of Microbisporicin Biosynthesis in Microbispora corallina

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