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.2013 Oct 29;4(6):e00498-13.
doi: 10.1128/mBio.00498-13.

Bactofencin A, a new type of cationic bacteriocin with unusual immunity

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Bactofencin A, a new type of cationic bacteriocin with unusual immunity

Eileen F O'Shea et al. mBio..

Abstract

Bacteriocin production is an important probiotic trait of intestinal bacteria. In this study, we identify a new type of bacteriocin, bactofencin A, produced by a porcine intestinal isolate Lactobacillus salivarius DPC6502, and assess its potency against pathogenic species including Staphylococcus aureus and Listeria monocytogenes. Genome sequencing of the bacteriocin producer revealed bfnA, which encodes the mature and highly basic (pI 10.59), 22-amino-acid defensin-like peptide. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectral analysis determined that bactofencin A has a molecular mass of 2,782 Da and contains two cysteine residues that form an intramolecular disulfide bond. Although an ABC transporter and transport accessory protein were also present within the bacteriocin gene cluster, a classical bacteriocin immunity gene was not detected. Interestingly, a dltB homologue was identified downstream of bfnA. DltB is usually encoded within the dlt operon of many Gram-positive bacteria. It is responsible for d-alanylation of teichoic acids in the cell wall and has previously been associated with bacterial resistance to cationic antimicrobial peptides. Heterologous expression of this gene conferred bactofencin A-specific immunity on sensitive strains of L. salivarius and S. aureus (although not L. monocytogenes), establishing its role in bacteriocin immunity. An analysis of the distribution of bfnA revealed that it was present in four additional isolates derived from porcine origin and absent from five human isolates, suggesting that its distribution is host specific. Given its novelty, we anticipate that bactofencin A represents the prototype of a new class of bacteriocins characterized as being cationic, with a DltB homologue providing a cognate immunity function.

Importance: This study describes the identification, purification, and characterization of bactofencin A, a novel type of bacteriocin produced by L. salivarius DPC6502. Interestingly, bactofencin A is not similar to any other known bacteriocin but instead shares similarity with eukaryotic cationic antimicrobial peptides, and here, we demonstrate that it inhibits two medically significant pathogens. Genome sequence analysis of the producing strain also revealed the presence of an atypical dltB homologue in the bacteriocin gene cluster, which was lacking a classical bacteriocin immunity gene. Furthermore, cloning this gene rendered sensitive strains resistant to the bacteriocin, thereby establishing its role in providing cognate bacteriocin immunity. Four additional L. salivarius isolates, also of porcine origin, were found to contain the bacteriocin biosynthesis genes and successfully produced bactofencin A, while these genes were absent from five human-derived strains investigated.

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Figures

FIG 1
FIG 1
HPLC profile (A), MALDI-TOF MS data (B), and antimicrobial activity (C) of purified bactofencin A. WVL, wavelength.
FIG 2
FIG 2
Nucleotide sequence and deduced peptide sequence of the structural gene of bactofencin A. The leader sequence is underlined, and the GG-processing site is indicated by an arrowhead. The bacteriocin structural gene and predicted immunity gene and transport genes are indicated by black, white, and gray arrows, respectively.
FIG 3
FIG 3
Inhibitory effect of synthetic bactofencin A on the growth of the indicator strainsL. monocytogenes NCTC 11994 (A) andS. aureus DPC5246 (B) at concentrations of 0 µM (gray diamonds), 0.05 µM (open circles), 0.1 µM (black circles), 0.5 µM (open squares), 1.0 µM (×), 5.0 µM (open trangles), and 10.0 µM (black squares). Error bars represent standard deviations based on triplicate data.
FIG 4
FIG 4
Viability ofS. aureus DPC5246 treated with (black squares) and without (open circles) synthetic bactofencin A as a function of time. These results represent the average of three independent experiments. Error bars represent standard errors based on triplicate data.
FIG 5
FIG 5
(A)S. aureus DPC5246 containing pEOS01 exhibits significantly enhanced resistance to bactofencin A relative to the resistance of the isogenic control strain at concentrations of 0.5 µM (P < 0.01), 2.5 µM (P < 0.05), and 5 µM (P < 0.05) following 24 h of incubation with the peptide. Error bars represent standard deviations based on triplicate data. (B)L. salivarius UCC118 containing pEOS01 (ii) also exhibits enhanced resistance to bactofencin A relative to the resistance of the respective isogenic control strain harboring pNZ44 (i).
FIG 6
FIG 6
The specific immunity ofL. salivarius UCC118 containing pEOS01 to bactofencin A (ii) (b) relative to the resistance ofL. salivarius UCC118 containing pNZ44 (i) (b) was not extended to the additional cationic antimicrobial peptides investigated, gramicidin (a), protegrin-1 (c), magainin II (d), (e) polymyxin B (e), ε-polylysine (f), and nisin (g).
FIG 7
FIG 7
Comparison of relative net cell surface charge. ThedltA-deficientS. aureus Sa113 experimental control has a higher affinity for cytochromec than wild-typeS. aureus Sa113 due to a decrease ind-alanylation of teichoic acids and a resultant increase in the net negative cell surface charge. However, the expression of BfnI does not influence the affinity ofS. aureus DPC5246 orL. salivarius UCC118 for cytochromec relative to the affinities of their respective controls.
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References

    1. Claesson MJ, van Sinderen D, O’Toole PW. 2008. Lactobacillus phylogenomics—towards a reclassification of the genus. Int. J. Syst. Evol. Microbiol. 58:2945–2954 - PubMed
    1. Wiedemann I, Böttiger T, Bonelli RR, Schneider T, Sahl HG, Martínez B. 2006. Lipid II-based antimicrobial activity of the lantibiotic plantaricin C. Appl. Environ. Microbiol. 72:2809–2814 - PMC - PubMed
    1. Joerger MC, Klaenhammer TR. 1986. Characterization and purification of helveticin J and evidence for a chromosomally determined bacteriocin produced by Lactobacillus helveticus 481. J. Bacteriol. 167:439–446 - PMC - PubMed
    1. Barrett E, Hayes M, O’Connor P, Gardiner G, Fitzgerald GF, Stanton C, Ross RP, Hill C. 2007. Salivaricin P: one of a family of two component anti-listerial bacteriocins produced by intestinal isolates of Lactobacillus salivarius. Appl. Environ. Microbiol. 73:3719–3723 - PMC - PubMed
    1. Dunne C, Murphy L, Flynn S, O’Mahony L, O’Halloran S, Feeney M, Morrissey D, Thornton G, Fitzgerald G, Daly C, Kiely B, Quigley EM, O’Sullivan GC, Shanahan F, Collins JK. 1999. Probiotics: from myth to reality. Demonstration of functionality in animal models of disease and in human clinical trials. Antonie Van Leeuwenhoek 76:279–292 - PubMed

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