Purification and structure of mutacin B-Ny266: a new lantibiotic produced by Streptococcus mutans

• Published in: FEBS letters 1997-06, Vol.410 (2), p.275-279
• Main Authors: Mota-Meira, Marilaine, Lacroix, Christophe, LaPointe, Gisèle, Lavoie, Marc C.
• Format: Article
• Language: English
• Subjects: 2,3-didehydroalanine; 2,3-didehydrobutyrine; 2-aminobutyric acid; Abu; American Type Culture Collection; Amino Acid Sequence; arbitrary units; ATCC; Bacteriocin; Bacteriocins - chemistry; Bacteriocins - isolation & purification; BM-Lan; BMSEC; dhA or U; dhB or O; DSER; high-pressure liquid chromatography; HPLC; Lan; lanthionine; Lantibiotic; Mass Spectrometry; Molecular Sequence Data; Mutacin; o-phthaldialdehyde; OPA; Organophosphorus Compounds - pharmacology; phenylthiohydantoin; PTH; PTH-dithiothreitol adduct of dhA; reversed-phase high-pressure liquid chromatography; RP-HPLC; S-ethylcysteine; SEC; Sequence Homology, Amino Acid; Streptococcus mutans; Streptococcus mutans - chemistry; Sulfhydryl Compounds - pharmacology; TFA; trifluoroacetic acid; trypticase soy agar yeast extract; TSAYE; β-methyl-S-ethylcysteine; β-methyllanthionine
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(97)00425-0

Mutacins are bactericidal substances of proteinaceous nature produced by Streptococcus mutans. Lantibiotics are antibacterial substances containing post-translationally modified amino acids such as lanthionine. Mutacin B-Ny266 was purified from the cell pellet of S. mutans strain Ny266 by ethanol extraction at pH 2.0 followed by reversed-phase chromatography (Sep-Pak® cartridge) and by HPLC on a C 18 column. The mean purification factor was 3240±81 and the mean yield was 1.0±0.1%. Molecular mass of mutacin B-Ny266 as determined by mass spectroscopy is 2270.29±0.21 Da. The amino acid sequence of the purified active fraction was obtained by Edman degradation after treatment with alkaline ethanethiol. Twenty-one amino acids were detected in this analysis. Mutacin B-Ny266 belongs to the type A lantibiotics. The proposed sequence is: F–K– A –W–U–F– A – Abu –P–G– A –A–K–O–G– A –F–N– U –Y– A . The molecule differs from that of epidermin/staphylococcin 1580 and gallidermin at positions 1, 2, 4, 5 and 6.