Isolation and biochemical characterization of a novel lantibiotic mutacin from Streptococcus mutans

Journal of Bacteriology - Tập 176 Số 14 - Trang 4316-4320 - 1994
Jan Novák1, Page W. Caufield1, Edward J. Miller1
1Department of Oral Biology, School of Dentistry, University of Alabama at Birmingham 35294.

Tóm tắt

Certain members of the indigenous biota of humans produce antimicrobial substances called bacteriocins, which inhibit other bacteria, including members of their own species. One of these substances, mutacin, is made by Streptococcus mutans, a member of the oral biota. Mutacin inhibits other mutans streptococci as well as many gram-positive exogenous pathogens. Here, we report for the first time the purification and partial biochemical characterization of a lanthionine-containing mutacin peptide from S. mutants T8. The biologically active peptide was isolated from the broth cultures by ultrafiltration and differential precipitation. The final mutacin preparation was homogeneous as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and N-terminal amino acid sequencing. A molecular mass of the peptide was estimated by electrospray ionization mass spectroscopy to be 3,244.64 +/- 1.15 Da. Its amino acid composition indicates the presence of lanthionine and likely beta-methyllanthionine in a total of about 25 amino acids. Because alpha,beta-unsaturated amino acids, the precursors of lanthionine residues, are often found in lantibiotics, we carried out the addition reaction of the mutacin with N-(methyl)mercaptoacetamide. The subsequent electrospray ionization mass spectroscopy analysis indicated the presence of two reaction products with M(r)s of 3,350.45 and 3,456.0. These are interpreted as the mutacin molecule with the addition of one and two molecules of reagent to the unsaturated amino acids, respectively. Sequencing of the peptide revealed an N-terminal amino acid sequence of Asn-Arg-Trp-Trp-Gln-Gly-Val-Val.

Từ khóa


Tài liệu tham khảo

Allgaier , H. , G. Jung , R. G. Werner , U. Schneider , and H. Zahner . 1985 . Elucidation of the structure of epidermin, a ribosomally synthesized tetracyclic heterodetic polypeptide antibiotic . Angew. Chem. 24 : 1051 - 1053 .

Allgaier , H. , G. Jung , R. G. Werner , U. Schneider , and H. Zahner . 1986 . Epidermin: sequencing of a heterodet tetracyclic 21-peptide amide antibiotic . Eur. J. Biochem. 160 : 9 - 22 .

Berkowitz , R. J. , and P. Jones . 1985 . Mouth-to-mouth transmission of the bacterium Streptococcus mutans between mother and child . Arch. Oral Biol. 30 : 377 - 379 .

Bradford , M. M. 1976 . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding . Anal. Biochem. 72 : 248 - 254 .

Buchman , G. W. , S. Banerje , and J. N. Hansen . 1988 . Structure, expression, and evolution of a gene encoding the precursor of nisin, a small protein antibiotic . J. Biol. Chem. 263 : 16260 - 16266 .

Caufield , P. W. , N. K. Childers , D. N. Allen , and J. B. Hansen . 1985 . Distinct bacteriocin groups correlate with different groups of Streptococcus mutans plasmids . Infect. Immun. 48 : 51 - 56 .

Caufield , P. W. , G. R. Shah , and S. K. Hollingshead . 1990 . Use of transposon Tn 916 to inactivate and isolate a mutacin-associated gene from Streptococcus mutans . Infect. Immun. 58 : 4126 - 4135 .

Caufield , P. W. , G. Shah , S. K. Hollingshead , M. Parrot , and M. C. Lavoie . 1990 . Evidence that mutacin II production is not mediated by a 5.6-kb plasmid in Streptococcus mutans . Plasmid 24 : 110 - 118 .

Computer Genetics Group. 1991. Program manual for the GCG package version 7. Computer Genetics Group Madison Wis.

Davey , A. L. , and A. H. Rogers . 1984 . Multiple types of the bacterium Streptococcus mutans in the human mouth and their intra-family transmission . Arch. Oral Biol. 29 : 453 - 460 .

Delisle , A. L. 1986 . Properties of mutacin b, an antibacterial substance produced by Streptococcus mutans strain BHT . Microbios 46 : 21 - 28 .

Gross , E. , and H. Kiltz . 1973 . The number and nature of a,junsaturated amino acids in subtilin . Biochem. Biophys. Res. Commun. 50 : 559 - 565 .

Gross , E. , and J. L. Morell . 1968 . The number and nature of oa,,-unsaturated amino acids in nisin . FEBS Lett. 2 : 61 - 64 .

Gross , E. , and J. L. Morell . 1971 . The structure of nisin . J. Am. Chem. Soc. 93 : 4634 - 4635 .

Hamada , S. , H. Imanishi , and T. Ooshima . 1986 . Isolation and mode of action of a cell-free bacteriocin (mutacin) from serotype g Streptococcus mutans MT3791 . Zentralbl. Bakteriol. Mikrobiol. Hyg. Ser. A 261 : 287 - 298 .

Hamada , S. , and T. Ooshima . 1975 . Inhibitory spectrum of a bacteriocin-like substance (mutacin) produced by some strains of Streptococcus mutans . J. Dent. Res. 54 : 140 - 145 .

Hamada , S. , and H. D. Slade . 1980 . Biology, immunology, and cariogenicity of Streptococcus mutans . Microbiol. Rev. 44 : 331 - 384 .

Hillman , J. D. , K. P. Johnson , and B. I. Yaphe . 1984 . Isolation of a Streptococcus mutans strain producing a novel bacteriocin . Infect. Immun. 44 : 141 - 144 .

Hurst , A. 1981 . Nisin. Adv. Appl. Microbiol. 27 : 85 - 123 .

Ikeda , T. , T. Iwanami , M. Hirasawa , C. Watanabe , J. R. McGhee , and T. Shiota . 1982 . Purification and certain properties of a bacteriocin from Streptococcus mutans . Infect. Immun. 35 : 861 - 868 .

Kaletta , C. , and K. -D. Entian . 1989 . Nisin, a peptide antibiotic: cloning and sequencing of the nisA gene and posttranslational processing of its peptide product . J. Bacteriol. 171 : 1597 - 1601 .

Kaletta , C. , K. -D. Entian , R. Kellner , G. Jung , M. Reis , and H. -G. Sahl . 1989 . Pep 5, a new lantibiotic: structural gene isolation and prepeptide sequence . Arch. Microbiol. 152 : 16 - 19 .

Kellner R. G. Jung and H.-G. Sahl. 1991. Structure elucidation of the tricyclic lantibiotic Pep 5 containing eight positively charged amino acids p. 141-158. In G. Jung and H. G. Sahl (ed.) Nisin and novel lantibiotics. ESCOM Science Publishers Leiden The Netherlands.

Kelstrup , J. , and R J . Gibbons . 1969 . Bacteriocins from human and rodent streptococci . Arch. Oral Biol. 14 : 251 - 258 .

Kurita , T. , and M. Hirasawa . 1988 . Biological and biochemical characterization of novel lipid-like antibacterial substances (mutalipocins) produced by Streptococcus mutans strain 32K . J. Gen. Microbiol. 134 : 213 - 220 .

Loyola , R J. P. , I. Morisaki , K. Kitamura , and S. Hamada . 1992 . Purification and properties of extracellular mutacin, a bacteriocin from Streptococcus sobrinus . J. Gen. Microbiol. 138 : 269 - 274 .

Miller , E. J. , D. K. Furuto , and A. J. Narkates . 1991 . Quantitation of type I, III, and V collagens in human tissue samples by high-performance liquid chromatography of selected cyanogen bromide peptides . Anal. Biochem. 196 : 54 - 60 .

Miller , E. J. , A. J. Narkates , and M. A. Niemann . 1990 . Amino acid analysis of collagen hydrolysates by reverse-phase high-performance liquid chromatography of 9-fluorenylmethyl chloroformate derivatives . Anal. Biochem. 190 : 92 - 97 .

Moore , S. 1963 . On the oxidation of cysteine as cysteic acid . J. Biol. Chem. 238 : 235 - 237 .

Parrot , M. , P. W. Caufield , and M. C. Lavoie . 1990 . Preliminary characterization of four bacteriocins from Streptococcus mutans . Can. J. Microbiol. 36 : 123 - 130 .

Paul , D. , and H. D. Slade . 1975 . Production and properties of an extracellular bacteriocin from Streptococcus mutans bacteriocidal for group A and other streptococci . Infect. Immun. 12 : 1375 - 1385 .

Rogers , A. H. 1972 . Effect of the medium on bacteriocin production among strains of Streptococcus mutans . Appl. Microbiol. 24 : 294 - 295 .

Rogers , A. H. 1976 . Bacteriocin patterns of strains belonging to various serotypes of Streptococcus mutans . Arch. Oral Biol. 21 : 243 - 249 .

Schager , H. , and G. Jagow . 1987 . Tricine sodium dodecyl sulfatepolyacrylamide gel electrophoresis for the separation of proteins in the range from 1-100 kDa . Anal. Biochem. 166 : 368 - 379 .

Schnell , N. , K. -D. Entian , U. Schneider , F. Gotz , H. Zahner , R. Kellner , and G. Jung . 1988 . Prepeptide sequence of epidermin, a ribosomally synthesized antibiotic with four sulfide-rings . Nature (London) 333 : 276 - 278 .

Tagg , J. R. 1991 . Studies of "BLIS-ful " oral N. Z. Dent. J. 87 : 14 - 16 .

Tagg , J. R. , N. L. Ragland , and N. P. Dickson . 1990 . A longitudinal study of Lancefield group A streptococcus acquisitions by a group of young Dunedin schoolchildren . N. Z. Med. J. 103 : 429 - 431 .

van de Run , I. , and R. E. Kessler . 1980 . Growth characteristics of group A streptococci in a new chemically defined medium . Infect. Immun. 27 : 444 448.

van der Meer , J. R. , J. Polman , M. M. Beerthuyzen , R. J. Siezen , 0. P. Kuipers , and W. M. de Vos . 1993 . Characterization of the Lactococcus lactis nisin A operon genes nisP, encoding a subtilisinlike serine protease involved in precursor processing, and nisR, encoding a regulatory protein involved in nisin biosynthesis . J. Bacteriol. 175 : 2578 - 2588 .

Thông tin
Thông tin xuất bản

Journal of Bacteriology

Tập 176 Số 14

4316-4320

Thông tin tác giả