title: Mechanistic dissection of the enzyme complexes involved in biosynthesis of lacticin 3147 and nisin.
abstract: The thioether rings in the lantibiotics lacticin 3147 and nisin are posttranslationally introduced by dehydration of serines and threonines, followed by coupling of these dehydrated residues to cysteines. The prepeptides of the two-component lantibiotic lacticin 3147, LtnA1 and LtnA2, are dehydrated and cyclized by two corresponding bifunctional enzymes, LtnM1 and LtnM2, and are subsequently processed and exported via one bifunctional enzyme, LtnT. In the nisin synthetase complex, the enzymes NisB, NisC, NisT, and NisP dehydrate, cyclize, export, and process prenisin, respectively. Here, we demonstrate that the combination of LtnM2 and LtnT can modify, process, and transport peptides entirely different from LtnA2 and that LtnT can process and transport unmodified LtnA2 and unrelated peptides. Furthermore, we demonstrate a higher extent of NisB-mediated dehydration in the absence of thioether rings. Thioether rings apparently inhibited dehydration, which implies alternating actions of NisB and NisC. Furthermore, certain (but not all) NisC-cyclized peptides were exported with higher efficiency as a result of their conformation. Taken together, these data provide further insight into the applicability of Lactococcus lactis strains containing lantibiotic enzymes for the design and production of modified peptides.
authors.affiliation: BiOMaDe Technology Foundation, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
authors.forename: Anneke
authors.initials: A
authors.lastname: Kuipers
authors.forename: Jenny
authors.initials: J
authors.lastname: Meijer-Wierenga
authors.forename: Rick
authors.initials: R
authors.lastname: Rink
authors.forename: Leon D
authors.initials: LD
authors.lastname: Kluskens
authors.forename: Gert N
authors.initials: GN
authors.lastname: Moll
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001430:Bacteriocins / Q000096:biosynthesis*; D004798:Enzymes / Q000235:genetics / Q000378:metabolism*; D013294:Lactococcus lactis / Q000201:enzymology*; D008958:Models, Molecular; D009561:Nisin / Q000096:biosynthesis*
chemical_list: D001426:Bacterial Proteins; D001430:Bacteriocins; D004798:Enzymes; C082508:lacticin 481; D009561:Nisin
references.citation: Biochemistry. 2005 Sep 27;44(38):12827-34
references.pmid: 16171398
references.citation: J Tongji Med Univ. 1995;15(4):193-7
references.pmid: 8731921
references.citation: Mol Microbiol. 2006 Jul;61(2):271-3
references.pmid: 16771845
references.citation: Appl Environ Microbiol. 2007 Sep;73(18):5809-16
references.pmid: 17660303
references.citation: Appl Environ Microbiol. 2006 Dec;72(12):7626-33
references.pmid: 17041158
references.citation: Appl Environ Microbiol. 2005 Jan;71(1):562-5
references.pmid: 15640237
references.citation: Biochemistry. 2004 Mar 23;43(11):3049-56
references.pmid: 15023056
references.citation: J Biol Chem. 2001 Jan 19;276(3):1772-9
references.pmid: 11038353
references.citation: Appl Environ Microbiol. 1993 Dec;59(12):4363-6
references.pmid: 16349136
references.citation: Eur J Biochem. 1995 Jun 15;230(3):827-53
references.pmid: 7601145
references.citation: Org Lett. 2007 Aug 16;9(17):3343-6
references.pmid: 17650008
references.citation: Biochemistry. 2007 Nov 13;46(45):13179-89
references.pmid: 17929939
references.citation: J Biol Chem. 2007 Jul 20;282(29):21169-75
references.pmid: 17513866
references.citation: J Biol Chem. 1999 Dec 31;274(53):37544-50
references.pmid: 10608807
references.citation: Science. 2006 Sep 15;313(5793):1636-7
references.pmid: 16973881
references.citation: FEMS Microbiol Lett. 1996 Oct 15;144(1):89-93
references.pmid: 8870256
references.citation: Science. 2006 Mar 10;311(5766):1464-7
references.pmid: 16527981
references.citation: J Bacteriol. 1993 May;175(9):2578-88
references.pmid: 8478324
references.citation: Appl Environ Microbiol. 2006 Jun;72(6):4492-6
references.pmid: 16751576
references.citation: J Mol Microbiol Biotechnol. 2007;13(4):235-42
references.pmid: 17827974
references.citation: Microbiology (Reading). 2002 Nov;148(Pt 11):3561-3568
references.pmid: 12427947
references.citation: Bioorg Med Chem Lett. 2008 May 15;18(10):3025-8
references.pmid: 18294843
references.citation: Methods Mol Biol. 1995;47:195-9
references.pmid: 7550735
references.citation: Chem Rev. 2005 Feb;105(2):633-84
references.pmid: 15700960
references.citation: Science. 2004 Jan 30;303(5658):679-81
references.pmid: 14752162
references.citation: J Bacteriol. 1997 Mar;179(5):1475-81
references.pmid: 9045802
references.citation: Microbiology (Reading). 2000 Sep;146 ( Pt 9):2147-2154
references.pmid: 10974102
references.citation: J Biol Chem. 2004 May 21;279(21):22176-82
references.pmid: 15044440
references.citation: Proc Natl Acad Sci U S A. 2005 Dec 20;102(51):18584-9
references.pmid: 16339304
references.citation: Biochemistry. 2005 Jun 21;44(24):8873-82
references.pmid: 15952794
references.citation: Mol Microbiol. 1995 Apr;16(2):229-40
references.pmid: 7565085
references.citation: J Mol Microbiol Biotechnol. 2007;13(4):226-34
references.pmid: 17827973
references.citation: Biochimie. 1988 Apr;70(4):559-66
references.pmid: 2844302
references.citation: Science. 1999 Dec 17;286(5448):2361-4
references.pmid: 10600751
references.citation: J Appl Microbiol. 2000 Aug;89(2):249-60
references.pmid: 10971756
references.citation: Appl Environ Microbiol. 1998 Feb;64(2):439-45
references.pmid: 9464377
references.citation: Mol Microbiol. 2006 Nov;62(3):735-47
references.pmid: 17076667
references.citation: Trends Biotechnol. 1997 Apr;15(4):135-40
references.pmid: 9131833
references.citation: Appl Microbiol. 1975 Jun;29(6):807-13
references.pmid: 16350018
references.citation: FEMS Microbiol Rev. 2001 May;25(3):285-308
references.pmid: 11348686
references.citation: Eur J Biochem. 1993 Aug 15;216(1):281-91
references.pmid: 7689965
references.citation: Appl Environ Microbiol. 2007 Mar;73(6):1792-6
references.pmid: 17261515
references.citation: Appl Environ Microbiol. 1996 Feb;62(2):612-9
references.pmid: 8593062
references.citation: J Am Chem Soc. 2006 Feb 8;128(5):1420-1
references.pmid: 16448091
references.citation: Appl Environ Microbiol. 1997 Nov;63(11):4252-60
references.pmid: 9361411
references.citation: Chem Biol. 2006 Oct;13(10):1109-17
references.pmid: 17052615
references.citation: Anal Biochem. 1987 Nov 1;166(2):368-79
references.pmid: 2449095
references.citation: J Biol Chem. 1996 May 24;271(21):12294-301
references.pmid: 8647829
references.citation: Nat Rev Drug Discov. 2006 Apr;5(4):321-32
references.pmid: 16531990
references.citation: Mol Microbiol. 2006 Jul;61(2):285-96
references.pmid: 16771847