Purification and partial characterization of a bacteriocin produced by Eikenella corrodens

• Published in: Journal of applied microbiology Vol. 104; no. 2; pp. 508 - 514
• Main Authors: Apolônio, A.C.M, Carvalho, M.A.R, Bemquerer, M.P, Santoro, M.M, Pinto, S.Q, Oliveira, J.S, Santos, K.V, Farias, L.M
• Format: Journal Article
• Language: English
• Published: Oxford, UK Oxford, UK : Blackwell Publishing Ltd 01-02-2008; Blackwell Publishing Ltd; Blackwell Science
• Subjects: Amino Acid Sequence; Antibiosis; bacteriocin; Bacteriocins - biosynthesis; Bacteriocins - genetics; Bacteriocins - isolation & purification; Biological and medical sciences; Buffers; Chromatography, High Pressure Liquid; corrodecin; Eikenella corrodens; Eikenella corrodens - metabolism; Fundamental and applied biological sciences. Psychology; Gram-Negative Bacterial Infections - microbiology; Humans; Hydrogen-Ion Concentration; Mass Spectrometry; Microbiology; Molecular Sequence Data; oral microbiota; Peptide Hydrolases - pharmacology; Peptostreptococcus - metabolism; periodontal disease; Periodontitis - microbiology; Sequence Analysis, Protein; Sequence Homology, Amino Acid; Solvents - pharmacology; Characterization; Periodontal disease; Purification; Applied microbiology; oral microbiota; Bacteriocin; Eikenella corrodons; Bacteria; Microflora; Eikenella corrodens; corrodecin
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/j.1365-2672.2007.03565.x

The purpose of this study was to purify and characterize a bacteriocin produced by Eikenella corrodens A32E2. Peptostreptococcus anaerobius ATCC27337 was used as indicator strain in antagonistic assays for bacteriocin-producing E. corrodens A32E2. Protein extraction was influenced by pH and buffer composition. The protein was active in the pH range 6-8. Inhibitory activity was lost by both heating and treatment with proteolytic enzymes and decreased with organic solvents. The substance is rather unstable but maintains 100% of its activity after being exposed to acetone and when stored at -70°C. The antagonistic substance was first precipitated by ammonium sulfate and further partially purified by Mono-Q FPLC and C-18 HPLC. Mass spectrometry analysis showed that the molecular mass was 23 625 Da, and the sequence obtained for the N-terminus was: Met-Asn-Phe-Asp-Glu-Lys-Val-Gly-Lys-Val-X-Phe-Lys-Val-Gly-Asp. The evidence presented in this study supports the idea that an antagonistic substance produced by E. corrodens A32E2 isolated from a periodontal diseased site is a novel bacteriocin, which we designate corrodecin. We anticipated that corrodecin might play an important role at the periodontal site. This compound could also be attractive in biotechnological applications as an interesting tool for oral ecosystem control.