Isolation and biochemical characterization of a novel lantibiotic mutacin from Streptococcus mutans.
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.
Novák
J
J
Department of Oral Biology, School of Dentistry, University of Alabama at Birmingham 35294.
Caufield
P W
PW
Miller
E J
EJ
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001430:Bacteriocins / Q000737:chemistry* / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013295:Streptococcus mutans; D013853:Thioacetamide / Q000031:analogs & derivatives
D000596:Amino Acids; D001430:Bacteriocins; D013853:Thioacetamide; C034455:N-methylmercaptoacetamide; C069066:mutacin II