Production, Purification and Chemical Properties of an Antistaphylococcal Agent Produced by Staphylococcus epidermidis

Sahl, Hans‐Georg; Brandis, H.

doi:10.1099/00221287-127-2-377

Cited by 78 publications

(86 citation statements)

References 19 publications

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“…In contrast, we could not observe a similar effect in the Pep 5 indicator organism Staphylococcus simulans 22 (formerly Staphylococcus cohnii 22) (2). However, autolysin activity could be demonstrated in the supernatant of a cell suspension of S. simulans treated with Pep 5, including at least two different autolysins which hydrolyzed the glycan chain and peptide bridges of the murein.…”

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confidence: 43%

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Autolytic system of Staphylococcus simulans 22: influence of cationic peptides on activity of N-acetylmuramoyl-L-alanine amidase

1987

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Pep 5 and nisin are cationic peptide antibiotics which in addition to their membrane-disruptive action induce autolysis in staphylococci. To investigate the mechanism of lysis induction, the influence of the peptides on the activity of the N-acetylmuramoyl-L-alanine amidase of Staphylococcus simulans 22 was studied. In experiments with isolated cell walls at low ionic strength, the amidase activity was stimulated by the addition of Pep 5 and nisin, as well as by polylysine, streptomycin, and mono-and divalent cations. The concentrations necessary for activation depended on the nature of the cation and rahged from 5 ,uM for poly-L-lysine (n = 17) to 150 mM for Na+ at a cell wail concentration of 100 ,ug of cell wails per ml. No effect was observed if the cell walls were devoid of polyanionic constituents. Kinetic data suggested that the amidase bound to the teichoic and teichuronic acids of the cell wall and was thereby inhibited. Cationic molecules reversed this inhibition, most likely by displacing the enzyme from the polyanions. If the concentrations of the larger peptides were high in relation to cell wall concentration, the activation turned into inhibition, presumably by interfering with the access of the enzyme to its substrate. These experiments demonstrate that the activity of the amidase is modulated by basic peptides in vitro and help to explain how Pep 5 and nisin may cause lysis of treated cells.

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confidence: 43%

“…Purification and activity testing of Pep 5 have been described previously (22). Pep 5 was additionally subjected to RP C18 high-pressure liquid chro-INFLUENCE OF BASIC PEPTIDES ON AUTOLYSIN ACTIVITY hydrolysates were determined with 2,4-dinitrofluorobenzene and by a modified Park-Johnson procedure, respectively (6).…”

Section: Methodsmentioning

confidence: 99%

Autolytic system of Staphylococcus simulans 22: influence of cationic peptides on activity of N-acetylmuramoyl-L-alanine amidase

1987

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“…Staphylococcus simulans 22 (formerly Staphylococcus cohnii 22; Institute for Medical Microbiology and Immunology of the University of Bonn, Bonn, Germany [21]) and Micrococcus luteus ATCC 4698 (American Type Culture Collection) were used as indicator strains. Unless indicated otherwise all experiments were performed with early-log-phase cells (A 600 , about 0.5) grown in half-concentrated Mueller-Hinton broth (Oxoid, Basingstoke, United Kingdom) at 37ЊC for S. simulans 22 and 30ЊC for M. luteus ATCC 4698.…”

Section: Methodsmentioning

confidence: 99%

Mode of action of the lantibiotic mersacidin: inhibition of peptidoglycan biosynthesis via a novel mechanism?

Brötz

Bierbaum

Markus

et al. 1995

Antimicrob Agents Chemother

131

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Mersacidin is an antibiotic peptide produced byThe term lantibiotics designates a group of lanthionine (and/or 3-methyllanthionine)-containing peptides with antibiotic activity against gram-positive bacteria (25). In contrast to peptide antibiotics like bacitracin or gramicidin S (10), lantibiotics are ribosomally synthesized and show unique structural properties, which arise from posttranslational modifications (for reviews, see references 3 and 9).With a molecular mass of 1,825 Da, mersacidin is the smallest lantibiotic known so far. Its propeptide template consists of 20 amino acids, 8 of which are modified to yield 4 thioether amino acids: 3 3-methyllanthionines and 1 2-aminovinyl-2-methylcysteine. This leads to the formation of four intramolecular heterocyclic rings. Additionally, the unusual amino acid ␣,␤-didehydroalanine is found. Mersacidin carries no net charge and has overall hydrophobic properties (5). It is produced by Bacillus sp. strain HIL Y-85,54728 and is active against several gram-positive bacteria such as streptococci, bacilli, and staphylococci including methicillin-resistant Staphylococcus aureus strains; enterococci are relatively insusceptible (4, 14). Although in vitro mersacidin is less active than vancomycin, teicoplanin, and daptomycin (17), its activity in vivo is at least comparable to that of vancomycin in the treatment of experimental staphylococcal infections (including those caused by methicillin-resistant S. aureus strains) in mice (4,14).In addition to its potential therapeutical value, mersacidin has been the subject of increased interest because of its position among the lantibiotics described so far, which have been classified into two groups (9). The type A lantibiotics (e.g., Pep5 and nisin) are screw-shaped, positively charged, amphipathic molecules which exert their primary bactericidal action by the formation of pores in the cytoplasmic membrane (20). The type B lantibiotics, as defined so far (e.g., duramycin and cinnamycin), possess a globular shape, carry no net charge or a negative charge, and are described as inhibitors of phospholipase A 2 or the angiotensin-converting enzyme (3). Mersacidin does not completely conform to either group because of atypical structural features (9, 11).Here, we report that it is also different regarding its mode of bactericidal activity. We show that mersacidin inhibits peptidoglycan biosynthesis, probably on the level of transglycosylation. Furthermore, we demonstrate that on the molecular level its mode of action differs from those of glycopeptide antibiotics like vancomycin or teicoplanin. This opens the possibility of using a new target for the antibiotic treatment of methicillinresistant S. aureus strains and makes mersacidin an interesting tool for the investigation of peptidoglycan biosynthesis of gram-positive bacteria. MATERIALS AND METHODSBacterial strains and growth conditions. Staphylococcus simulans 22 (formerly Staphylococcus cohnii 22; Institute for Medical Microbiology and Immunology of the University of Bonn, Bonn, ...

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“…The mature peptide displays several unusual features, such as the dehydrated residues dehydroalanine and dehydrobutyrine, which are derived from Ser and Thr residues, respectively (22, 24), and lanthionine and P-methyllanthionine residues, which form five intramolecular thioether bridges. Molecular structures similar to those in mature nisin are found in several other antibacterial peptides secreted, e.g., by Bacillus subtilis (subtilin) (21) or Staphylococcus epidermidis (epidermin and Pep5) (2,42), together forming the group of the lantibiotics (47; for a review, see reference 25).…”

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confidence: 99%

Characterization of the Lactococcus lactis nisin A operon genes nisP, encoding a subtilisin-like serine protease involved in precursor processing, and nisR, encoding a regulatory protein involved in nisin biosynthesis

et al. 1993

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Biosynthesis of the lantibiotic peptide nisin by Lactococcus lactis NIZO R5 relies on the presence of the conjugative transposon TnS276 in the chromosome. A 12-kb DNA fragment of T M276 including the nisA gene and about 10 kb of downstream DNA was cloned in L. lactis, resulting in the production of an extracellular nisin precursor peptide. This peptide reacted with antibodies against either nisin A or the synthetic leader peptide, suggesting that it consisted of a fully modified nisin with the nisin leader sequence still attached to it. This structure was confirmed by N-terminal sequencing and 'H-nuclear magnetic resoqnnce analysis of the purified peptide. Deletion studies showed that the nisR gene is essential for the production of this intermediate. The deduced amino acid sequence of the nisR gene product indicated that the protein belongs to the family of two-component regulators. The deduced amino acid sequence of NisP, the putative product of the gene upstream of nisR, showed an N-terminal signal sequence, a catalytic domain with a high degree of similarity to those of subtilisin-like serine proteases, and a putative C-terminal membrane anchor. Cell extracts of Escherichia coli overexpressing nisP were able to cleave the nisin precursor peptide, producing active, mature nisin. A similar activation was obtained with whole cells but not with membrane-free extracts ofL. lactis strains carrying TnS276 in which the nisA gene had been inactivated. The results indicate that the penultimate step in nisin biosynthesis is secretion of precursor nisin without cleavage of the leader peptide, whereas the last step is the cleavage of the leader peptide sequence from the fully maturated nisin peptide.Nisin is a 34-residue antibacterial peptide that is produced by several strains of Lactococcus lactis and strongly inhibits the growth of a broad range of gram-positive bacteria (24). Insertion of the peptide into the cytoplasmic membrane of the target cell renders the membrane permeable to monovalent cations (18) and dissipates the proton motive force of the cell (18, 37). The mature peptide displays several unusual features, such as the dehydrated residues dehydroalanine and dehydrobutyrine, which are derived from Ser and Thr residues, respectively (22, 24), and lanthionine and P-methyllanthionine residues, which form five intramolecular thioether bridges. Molecular structures similar to those in mature nisin are found in several other antibacterial peptides secreted, e.g., by Bacillus subtilis (subtilin) (21) or Staphylococcus epidermidis (epidermin and Pep5) (2, 42), together forming the group of the lantibiotics (47; for a review, see reference 25).Recently, it was reported that two widely distributed natural variants of nisin exist, named nisin A and nisin Z, which differ in a single amino acid residue (11,36). The structural genes for both nisin A, nisA, and nisin Z, nisZ, have been cloned and sequenced elsewhere (5,14,26,36).These studies showed that both nisin variants are ribosomally synthesized as 57-amino-acid pre...

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Production, Purification and Chemical Properties of an Antistaphylococcal Agent Produced by Staphylococcus epidermidis

Cited by 78 publications

References 19 publications

Autolytic system of Staphylococcus simulans 22: influence of cationic peptides on activity of N-acetylmuramoyl-L-alanine amidase

Autolytic system of Staphylococcus simulans 22: influence of cationic peptides on activity of N-acetylmuramoyl-L-alanine amidase

Mode of action of the lantibiotic mersacidin: inhibition of peptidoglycan biosynthesis via a novel mechanism?

Characterization of the Lactococcus lactis nisin A operon genes nisP, encoding a subtilisin-like serine protease involved in precursor processing, and nisR, encoding a regulatory protein involved in nisin biosynthesis

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