incubated at 30 1C. In order to further study the reduction step,
the same assay was used except NaHCO3 was omitted. The assay
system are analyzed by HPLC-ESIMS with a flow rate of
1 mL minꢁ1 over a 30 min gradient as follows T = 0, 5% B;
T = 30, 40% B (A: 10 mM NH4OAc in water; B: acetonitrile).
(2S)-2-ethylmalonyl-CoA (M + H+: 881.90) and butyryl-CoA
(M + H+: 837.86) were eluted at 6.6 min and 9.0 min, respectively.
17 C. D. Reeves, S. Murli, G. W. Ashley, M. Piagentini, C. R. Hutchinson
and R. McDaniel, Biochemistry, 2001, 40, 15464–15470.
18 F. Del Vecchio, H. Petkovic, S. G. Kendrew, L. Low, B. Wilkinson,
R. Lill, J. Cortes, B. A. M. Rudd, J. Staunton and P. F. Leadlay,
´
J. Ind. Microbiol. Biotechnol., 2003, 30, 489–494.
19 H. Petkovic, R. E. Lill, R. M. Sheridan, B. Wilkinson,
E. L. McCormick, H. A. McArthur, J. Staunton, P. F. Leadlay
and S. G. Kendrew, J. Antibiot., 2003, 56, 543–551.
20 R. Reid, M. Piagentini, E. Rodriguez, G. Ashley, N. Viswanathan,
J. Carney, D. V. Santi, C. R. Hutchinson and R. McDaniel,
Biochemistry, 2003, 42, 72–79.
Nucleotide sequence accession number
21 P. Caffrey, ChemBioChem, 2003, 4, 654–657.
22 A. T. Keatinge-Clay, Chem. Biol., 2007, 14, 898–908.
23 D. J. Bevitt, J. Cortes, S. F. Haydock and P. F. Leadlay, Eur. J.
Biochem., 1992, 204, 39–49.
The nucleotide sequence reported in this study is available in
the GenBank database under accession no. FJ809786.
24 X. Ruan, D. Stassi, S. A. Lax and L. Katz, Gene, 1997, 203, 1–9.
25 P. F. Fitzpatrick, J. Am. Chem. Soc., 1994, 116, 1133–1134.
26 W. Zhang, B. Ostash and C. T. Walsh, Proc. Natl. Acad. Sci.
U. S. A., 2010, 107, 16828–16833.
27 D. G. Fujimori, S. Hrvatin, C. S. Neumann, M. Strieker,
M. A. Marahiel and C. T. Walsh, Proc. Natl. Acad. Sci. U. S. A.,
2007, 104, 16498–16503.
28 T. Stachelhaus, H. D. Mootz and M. A. Marahiel, Chem. Biol.,
1999, 6, 493–505.
29 G. L. Challis, J. Ravel and C. A. Townsend, Chem. Biol., 2000, 7,
211–224.
30 C. Rausch, T. Weber, O. Kohlbacher, W. Wohlleben and
D. H. Huson, Nucleic Acids Res., 2005, 33, 5799–5808.
31 L. Du and L. Lou, Nat. Prod. Rep., 2010, 27, 255–278.
32 T. J. Erb, V. Brecht, G. Fuchs, M. Muller and B. E. Alber, Proc.
Natl. Acad. Sci. U. S. A., 2009, 106, 8871–8876.
Acknowledgements
This work was supported in part by grants from NNSF
(30900021, 20902102, 20832009, 90713012 and 20921091),
MST (2009ZX09501-008), ‘‘973 program’’ (2010CB833200),
CAS (KJCX2-YW-H08 and KSCX2-YW-G-06), and STCSM
(09QH1402700) of China. We thank Nigel Coates, Matthew
Gregory, Teresa Foster, Steven Moss, Mahmood Piraee, Tony
Warneck and Ming Zhang at Biotica Technology Ltd. for help
with fermentation methods and analysis of strains.
References
33 T. J. Erb, J. Retey, G. Fuchs and B. E. Alber, J. Biol. Chem., 2008,
283, 32283–32293.
34 W. Zhang, B. D. Ames, S. Tsai and Y. Tang, Appl. Environ.
Microbiol., 2006, 72, 2573–2580.
35 Y. Chan, A. Podevels, B. Kevany and M. Thomas, Nat. Prod. Rep.,
2009, 26, 90–114.
36 H. Ikeda and S. Omura, Chem. Rev., 1997, 97, 2591–2610.
37 K. C. Nicolaou, J. Xu, F. Murphy, S. Barluenga, O. Baudoin,
H. Wei, D. L. Gray and T. Ohshima, Angew. Chem., Int. Ed., 1999,
38, 2447–2451.
38 L. A. Paquette, M. Duan, I. Konetzki and C. Kempmann, J. Am.
Chem. Soc., 2002, 124, 4257–4270.
39 L. A. Kelley and M. J. E. Sternberg, Nat. Protoc., 2009, 4, 363–371.
40 L. Falquet, M. Pagni, P. Bucher, N. Hulo, C. J. A. Sigrist,
K. Hofmann and A. Bairoch, Nucleic Acids Res., 2002, 30,
235–238.
1 J. J. Sanglier, V. Quesniaux, T. Fehr, H. Hofmann, M. Mahnke,
K. Memmert, W. Schuler, G. Zenke, L. Gschwind, C. Maurer and
W. Schilling, J. Antibiot., 1999, 52, 466–473.
2 B. Weidmann, K. Zimmermann and K. Lin, Intern. Patent. Appl.,
WO/2006/138507 , 2006.
3 C. Hartel, P. Iblher, A. Puzik, K. Wortmeier, B. Ebel, C. Schultz
and M. Muller-Steinhardt, Scand. J. Immunol., 2006, 63, 26–34.
4 E. Sokolskaja, D. M. Sayah and J. Luban, J. Virol., 2004, 78,
12800–12808.
5 S. Javadov, M. Karmazyn and N. Escobales, J. Pharmacol. Exp.
Ther., 2009, 330, 670–678.
6 J. Kallen, R. Sedrani, G. Zenke and J. Wagner, J. Biol. Chem.,
2005, 280, 21965–21971.
7 M. T. Ivery, Med. Res. Rev., 2000, 20, 452–484.
8 C. Steinschulte, T. Taner, A. W. Thomson, G. Bein and
H. Hackstein, J. Immunol., 2003, 171, 542–546.
41 W. Liu, S. D. Christenson, S. Standage and B. Shen, Science, 2002,
297, 1170–1173.
42 T. Kieser, M. Bibb, M. Butter, K. F. Chater and D. A. Hopwood,
Practical Streptomyces Genetics, The John Innes Foundation,
Norwich, 2001.
9 L. W. Miller, Circulation, 1996, 94, 1209–1211.
10 R. McDaniel, M. Welch and C. R. Hutchinson, Chem. Rev., 2005,
105, 543–558.
11 X. Jia, Z. Tian, L. Shao, X. Qu, Q. Zhao, J. Tang, G. Tang and
W. Liu, Chem. Biol., 2006, 13, 575–585.
43 J. Sambrook and D. W. Russell, Molecular Cloning: A Laboratory
Manual, Cold Spring Harbor Laboratory Press, New York,
3rd edn, 2001.
44 B. Gust, T. Kieser and K. F. Chater, REDIRECT Technology:
PCR Targeting System in STREPTOMYCES Coelicolor, John
Innes Centre, Norwich Research Park, Colney, Norwich, NR4
7UH, United Kingdom, 2002.
45 J. E. Coligan, B. M. Dunn, H. L. Ploegh, D. W. Speicher and
P. T. Wingfield, Current Protocols in Protein Science, John Wiley
and Sons, Inc., New York, 1995.
12 L. Li, W. Deng, J. Song, W. Ding, Q. Zhao, C. Peng, W. Song,
G. Tang and W. Liu, J. Bacteriol., 2008, 190, 251–263.
13 T. J. Erb, I. A. Berg, V. Brecht, M. Muller, G. Fuchs and B. E. Alber,
Proc. Natl. Acad. Sci. U. S. A., 2007, 104, 10631–10636.
14 Y. Liu, C. Hazzard, A. S. Eustaquio, K. A. Reynolds and
B. S. Moore, J. Am. Chem. Soc., 2009, 131, 10376–10377.
15 H. Liu and K. A. Reynolds, J. Bacteriol., 1999, 181, 6806–6813.
16 F. H. Stephen, F. A. Jesus, M. Istvan, S. Torsten, K. Lake Ee,
K. Ariane, F. A. M. Andrew, S. G. Ian, S. James and F. L. Peter,
FEBS Lett., 1995, 374, 246–248.
c
This journal is The Royal Society of Chemistry 2011
Mol. BioSyst., 2011, 7, 852–861 861