title: Production of the novel two-peptide lantibiotic lichenicidin by Bacillus licheniformis DSM 13.
abstract: BACKGROUND
  Lantibiotics are small microbial peptide antibiotics that are characterized by the presence of the thioether amino acids lanthionine and methyllanthionine. Lantibiotics possess structural genes which encode inactive prepeptides. During maturation, the prepeptide undergoes posttranslational modifications including the introduction of rare amino acids as lanthionine and methyllanthione as well as the proteolytic removal of the leader. The structural gene (lanA) as well as the other genes which are involved in lantibiotic modification (lanM, lanB, lanC, lanP), regulation (lanR, lanK), export (lanT(P)) and immunity (lanEFG) are organized in biosynthetic gene clusters.
  
  
  RESULTS
  Sequence comparisons in the NCBI database showed that Bacillus licheniformis DSM 13 harbours a putative lantibiotic gene cluster which comprises two structural genes (licA1, licA2) and two modification enzymes (licM1, licM2) in addition to 10 ORFs that show sequence similarities to proteins involved in lantibiotic production. A heat labile antimicrobial activity was detected in the culture supernatant and a heat stabile activity was present in the isopropanol cell wash extract of this strain. In agar well diffusion assays both fractions exhibited slightly different activity spectra against Gram-positive bacteria. In order to demonstrate the connection between the lantibiotic gene cluster and one of the antibacterial activities, two Bacillus licheniformis DSM 13 mutant strains harbouring insertions in the structural genes of the modification enzymes licM1 and licM2 were constructed. These strains were characterized by a loss of activity in the isopropanol extract and substractive MALDI-TOF predicted masses of 3020.6 Da and 3250.6 Da for the active peptides.
  
  
  CONCLUSIONS
  In conclusion, B. licheniformis DSM 13 produces an antimicrobial substance that represents the two-peptide lantibiotic lichenicidin and that shows activity against a wide range of Gram-positive bacteria including methicillin resistant Staphylococcus aureus strains.
authors.affiliation: Institute of Medical Microbiology, Immunology and Parasitology, University of Bonn, Bonn, Germany.
authors.forename: Jasmin
authors.initials: J
authors.lastname: Dischinger
authors.forename: Michaele
authors.initials: M
authors.lastname: Josten
authors.forename: Christiane
authors.initials: C
authors.lastname: Szekat
authors.forename: Hans-Georg
authors.initials: HG
authors.lastname: Sahl
authors.forename: Gabriele
authors.initials: G
authors.lastname: Bierbaum
mesh_terms: D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology; D001407:Bacillus / Q000235:genetics / Q000378:metabolism*; D001430:Bacteriocins / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology; D005798:Genes, Bacterial; D055624:Methicillin-Resistant Staphylococcus aureus / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D005810:Multigene Family; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000900:Anti-Bacterial Agents; D001430:Bacteriocins; C546394:lichenicidin, Bacillus licheniformis
references.citation: Annu Rev Microbiol. 2007;61:477-501
references.pmid: 17506681
references.citation: Appl Environ Microbiol. 1995 May;61(5):1706-13
references.pmid: 7646007
references.citation: Appl Environ Microbiol. 2004 Nov;70(11):6887-91
references.pmid: 15528558
references.citation: Proc Natl Acad Sci U S A. 2003 Apr 1;100(7):4316-21
references.pmid: 12642677
references.citation: FEMS Microbiol Lett. 1995 Mar 15;127(1-2):121-6
references.pmid: 7737474
references.citation: Chem Biol. 2008 Oct 20;15(10):1035-45
references.pmid: 18940665
references.citation: Appl Microbiol Biotechnol. 2008 Feb;78(1):181-8
references.pmid: 18046550
references.citation: Eur J Biochem. 2000 Jul;267(13):4068-74
references.pmid: 10866808
references.citation: J Biol Chem. 2001 Jan 19;276(3):1772-9
references.pmid: 11038353
references.citation: Antimicrob Agents Chemother. 1985 May;27(5):836-40
references.pmid: 4015073
references.citation: Mol Microbiol. 1998 Aug;29(4):1029-38
references.pmid: 9767571
references.citation: Antimicrob Agents Chemother. 2006 Apr;50(4):1449-57
references.pmid: 16569864
references.citation: Biochemistry. 2007 May 22;46(20):5884-95
references.pmid: 17469849
references.citation: Mini Rev Med Chem. 2007 Dec;7(12):1236-47
references.pmid: 18220976
references.citation: Mol Microbiol. 2005 Dec;58(5):1368-80
references.pmid: 16313622
references.citation: J Antibiot (Tokyo). 1992 Jun;45(6):832-8
references.pmid: 1500347
references.citation: Appl Environ Microbiol. 1998 Dec;64(12):4803-8
references.pmid: 9835565
references.citation: Curr Pharm Biotechnol. 2009 Jan;10(1):2-18
references.pmid: 19149587
references.citation: J Biol Chem. 1999 Dec 31;274(53):37544-50
references.pmid: 10608807
references.citation: Genome Biol. 2004;5(10):R77
references.pmid: 15461803
references.citation: J Bacteriol. 1987 Dec;169(12):5452-8
references.pmid: 2890620
references.citation: J Mol Microbiol Biotechnol. 2004;7(4):204-11
references.pmid: 15383718
references.citation: Eur J Biochem. 1988 Oct 15;177(1):53-9
references.pmid: 3181159
references.citation: Proc Natl Acad Sci U S A. 2006 Nov 14;103(46):17243-8
references.pmid: 17085596
references.citation: Nucleic Acids Res. 2000 Nov 1;28(21):4317-31
references.pmid: 11058132
references.citation: Infect Immun. 1999 Aug;67(8):4268-71
references.pmid: 10417203
references.citation: Mol Microbiol. 2006 Feb;59(3):731-42
references.pmid: 16420347
references.citation: Antimicrob Agents Chemother. 2005 Jul;49(7):2606-11
references.pmid: 15980326
references.citation: Int J Med Microbiol. 2009 Jun;299(5):313-22
references.pmid: 19138560
references.citation: Appl Environ Microbiol. 2006 Nov;72(11):7270-7
references.pmid: 16980420
references.citation: Antimicrob Agents Chemother. 1998 Jan;42(1):154-60
references.pmid: 9449277
references.citation: Proc Natl Acad Sci U S A. 2004 Aug 3;101(31):11448-53
references.pmid: 15277670
references.citation: J Bacteriol. 1994 Dec;176(23):7335-44
references.pmid: 7961506
references.citation: Appl Environ Microbiol. 1996 Feb;62(2):385-92
references.pmid: 8593044
references.citation: FEMS Microbiol Lett. 2007 Feb;267(1):64-71
references.pmid: 17233677
references.citation: Curr Protein Pept Sci. 2005 Feb;6(1):77-84
references.pmid: 15638770
references.citation: J Bacteriol. 1985 May;162(2):833-6
references.pmid: 3921529
references.citation: Biochemistry. 2004 Mar 23;43(11):3049-56
references.pmid: 15023056
references.citation: FEMS Microbiol Lett. 2005 Nov 15;252(2):235-41
references.pmid: 16194596
references.citation: Mol Microbiol. 1995 Jun;16(5):969-76
references.pmid: 7476193
references.citation: Eur J Biochem. 1995 Sep 1;232(2):478-89
references.pmid: 7556197
references.citation: Biotechnol Prog. 1997 Nov-Dec;13(6):757-61
references.pmid: 9413133
references.citation: Appl Environ Microbiol. 2000 Jun;66(6):2565-71
references.pmid: 10831439
references.citation: Mol Microbiol. 2006 Nov;62(3):735-47
references.pmid: 17076667
references.citation: Eur J Biochem. 1986 Oct 1;160(1):9-22
references.pmid: 3769923
references.citation: Acta Microbiol Pol A. 1976;8(1):71-7
references.pmid: 820164
references.citation: Microbiology (Reading). 2001 Mar;147(Pt 3):643-651
references.pmid: 11238971
references.citation: N Engl J Med. 1988 Jul 21;319(3):157-61
references.pmid: 2968517
references.citation: Antimicrob Agents Chemother. 2005 Feb;49(2):541-8
references.pmid: 15673730
references.citation: Mol Microbiol. 2006 Jul;61(2):285-96
references.pmid: 16771847
references.citation: Microbiology (Reading). 2003 Jan;149(Pt 1):9-17
references.pmid: 12576575
references.citation: Appl Microbiol. 1971 May;21(5):943
references.pmid: 4930039