Mechanistic dissection of the enzyme complexes involved in biosynthesis of lacticin 3147 and nisin.
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.
Kuipers
Anneke
A
BiOMaDe Technology Foundation, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
Meijer-Wierenga
Jenny
J
Rink
Rick
R
Kluskens
Leon D
LD
Moll
Gert N
GN
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*
D001426:Bacterial Proteins; D001430:Bacteriocins; D004798:Enzymes; C082508:lacticin 481; D009561:Nisin
Biochemistry. 2005 Sep 27;44(38):12827-34
16171398
J Tongji Med Univ. 1995;15(4):193-7
8731921
Mol Microbiol. 2006 Jul;61(2):271-3
16771845
Appl Environ Microbiol. 2007 Sep;73(18):5809-16
17660303
Appl Environ Microbiol. 2006 Dec;72(12):7626-33
17041158
Appl Environ Microbiol. 2005 Jan;71(1):562-5
15640237
Biochemistry. 2004 Mar 23;43(11):3049-56
15023056
J Biol Chem. 2001 Jan 19;276(3):1772-9
11038353
Appl Environ Microbiol. 1993 Dec;59(12):4363-6
16349136
Eur J Biochem. 1995 Jun 15;230(3):827-53
7601145
Org Lett. 2007 Aug 16;9(17):3343-6
17650008
Biochemistry. 2007 Nov 13;46(45):13179-89
17929939
J Biol Chem. 2007 Jul 20;282(29):21169-75
17513866
J Biol Chem. 1999 Dec 31;274(53):37544-50
10608807
Science. 2006 Sep 15;313(5793):1636-7
16973881
FEMS Microbiol Lett. 1996 Oct 15;144(1):89-93
8870256
Science. 2006 Mar 10;311(5766):1464-7
16527981
J Bacteriol. 1993 May;175(9):2578-88
8478324
Appl Environ Microbiol. 2006 Jun;72(6):4492-6
16751576
J Mol Microbiol Biotechnol. 2007;13(4):235-42
17827974
Microbiology (Reading). 2002 Nov;148(Pt 11):3561-3568
12427947
Bioorg Med Chem Lett. 2008 May 15;18(10):3025-8
18294843
Methods Mol Biol. 1995;47:195-9
7550735
Chem Rev. 2005 Feb;105(2):633-84
15700960
Science. 2004 Jan 30;303(5658):679-81
14752162
J Bacteriol. 1997 Mar;179(5):1475-81
9045802
Microbiology (Reading). 2000 Sep;146 ( Pt 9):2147-2154
10974102
J Biol Chem. 2004 May 21;279(21):22176-82
15044440
Proc Natl Acad Sci U S A. 2005 Dec 20;102(51):18584-9
16339304
Biochemistry. 2005 Jun 21;44(24):8873-82
15952794
Mol Microbiol. 1995 Apr;16(2):229-40
7565085
J Mol Microbiol Biotechnol. 2007;13(4):226-34
17827973
Biochimie. 1988 Apr;70(4):559-66
2844302
Science. 1999 Dec 17;286(5448):2361-4
10600751
J Appl Microbiol. 2000 Aug;89(2):249-60
10971756
Appl Environ Microbiol. 1998 Feb;64(2):439-45
9464377
Mol Microbiol. 2006 Nov;62(3):735-47
17076667
Trends Biotechnol. 1997 Apr;15(4):135-40
9131833
Appl Microbiol. 1975 Jun;29(6):807-13
16350018
FEMS Microbiol Rev. 2001 May;25(3):285-308
11348686
Eur J Biochem. 1993 Aug 15;216(1):281-91
7689965
Appl Environ Microbiol. 2007 Mar;73(6):1792-6
17261515
Appl Environ Microbiol. 1996 Feb;62(2):612-9
8593062
J Am Chem Soc. 2006 Feb 8;128(5):1420-1
16448091
Appl Environ Microbiol. 1997 Nov;63(11):4252-60
9361411
Chem Biol. 2006 Oct;13(10):1109-17
17052615
Anal Biochem. 1987 Nov 1;166(2):368-79
2449095
J Biol Chem. 1996 May 24;271(21):12294-301
8647829
Nat Rev Drug Discov. 2006 Apr;5(4):321-32
16531990
Mol Microbiol. 2006 Jul;61(2):285-96
16771847