Shen Lab Research: Project 2

(ii)   Discovery and elucidation of novel enzymes, biochemistry, and mechanism of catalysis

1. Discovery and biochemical characterization of BlmI as the first type II peptidyl carrier protein (PCP) (Chem. Biol. 1999, 6, 507-517)

2. Proposal of the bleomycin biosynthetic gene cluster as a model hybrid peptide synthetase (NRPS)-polyketide synthase (PKS) for hybrid peptide-polyketide biosynthesis (Chem. Biol. 2000, 7, 623-642; Met. Engineer. 2001, 3, 78-95; Curr. Opin. Drug Disco. Decelop. 2001, 4, 215-228; J. Ind. Microbiol. Biotechnol. 2001, 27, 378-385; J. Nat. Prod. 2002, 65, 422-531)

3. Discovery of an oxidation (Ox) domain of NRPS for thiazole biosynthesis (FEMS Microbiol. Lett. 2000, 189, 171-175; Biochemistry 2003, 42, 9722-9730)

4. Characterization of triple hydroxylation by TcmG or ElmG (J. Biol. Chem. 1994, 48, 30726-30733; Org. Lett. 2000, 2, 3225-3227; J. Nat. Prod. 2001, 64, 444-449)

5. Discovery of a pair of enantio-complementary pathways for macrotetrolide biosynthesis (Antimicrob. Agents Chemther. 2000, 44, 1943-1953; J. Am. Chem. Soc. 2001, 123, 3385-3386)

6. Discovery of the Svp phosphopantetheinyl transferase with broad substrate specificity for both ACP and PCP to activate hybrid NRPS-PKS (Chem. Biol. 2001, 8, 725-738)

7. Discovery of an iterative PKS for enediyne biosynthesis (Science 2002, 297, 1170-1173; Nat. Biotechnol. 2003, 21, 187-190)

8. Discovery of a type II PKS for C-O bond formation (Science 2002, 297, 1327-1330; Chem. Rec. 2002, 2, 389-396)

9. Development of a PCR method for NRPS domains (Cy and Ox domains) to clone thiazole-containing natural product biosynthetic pathways (J. Bacteriol. 2002, 184, 7013-7024)

10. Discovery of NRPS domain “skipping” (A and Ox domains) in nonribosomal peptide biosynthesis (Biochemistry 2003, 42, 9722-9730; Biochemistry 2003, 42, 9731-9740)

11. Discovery of an AT-less PKS (Proc. Natl. Acad. Sci. USA 2003, 100, 3149-3154; Chem. Biol. 2004, 11, 33-45)

12. Discovery of a 4-methylideneimidazole-5-one (MIO)-containing tyrosine aminomutase (J. Am. Chem. Soc. 2003, 125, 6062-6063; Biochemistry 2003, 42, 12708-12718; J. Am. Chem. Soc. 2007, 129, 15744-15745; Bioorg. Med. Chem. Lett. 2008, 18, 3099-3102)

13. Determination of the structure of the SgcC4 4-methylideneimidazole-5-one (MIO)-containing tyrosine aminomutase (Biochemistry 2007, 46, 7205-7214)

14. Development of a PCR method for enediyne PKS to clone enediyne biosynthetic pathways (Proc. Natl. Acad. Soc. USA  2003, 100, 11959-11963)

15. Characterization of SgcA1 as a a-D-glucopyranosyl-1-phosphate thymidylytransferase for biosynthesis of the C-1027 enediyne antitumor antibiotic (J. Nat. Prod. 2004, 67, 206-213)

16. Discovery of migrastatin and dorrigocins as shunt metabolites of iso-migrastatin (J. Am. Chem. Soc. 2005, 127, 1622-1623)

17. Discovery of an adenylation enzyme (A) of NRPS that activate b-tyrosine (J. Am. Chem. Soc. 2005, 127, 11594-11595; J. Biol. Chem. 2006, 281, 29633-29640)

18. Discovery of PKS domain “skipping” (ACP domain) in polyketide biosynthesis by type I PKS (J. Nat. Prod. 2006, 69, 387-393)

19. Development of a PCR method for cloning methoxymalonate-incorporating polyketide biosynthetic pathways (J. Bacteriol. 2006, 198, 4148-4152; J. Bacteriol. 2006, 198, 4141-4147)

20. Discovery of the first holo-ACP synthase-type phosphopantetheinyl transferase for a type II PKS (J. Am. Chem. Soc. 2005, 127, 16442-16452; J. Biol. Chem. 2006, 281, 29660-29668)

21. Identification of 1,3-bisphosphoglycerate as the direct precursor for methoxymalonyl-ACP biosynthesis (J. Am. Chem. Soc. 2006, 128, 10386-10387)

22. Discovery of a [3,3]-sigmtropic rearrangement for iso-migratsatin to migratsatin (Org. Lett. 2006, 8, 5865-5868)

23. Characterization of NcsB2 as a promiscuous naphthoic acid-coenzyme A ligase for the biosynthesis of the neocarzinostatin enediyne antitumor antibiotic (J. Am. Chem. Soc. 2007, 129, 7728-7729)

24. Discovery of an unprecedented NRPS initiation module that is consisted of a discrete D-alanine-specific adenylation enzyme (LnmQ) and a discrete PCP (LnmP) (J. Biol. Chem. 2007, 282, 20273-20282)

25. Characterization of SgcC3 as an FAD-dependent halogenase that acts on a carrier protein-tethered substrate for the biosynthesis of the C-1027 enediyne antitumor antibiotic (J. Am. Chem. Soc. 2007, 129, 12432-12438)

26. Discovery of a new branch point in chorismate metabolism from the biosynthesis of the C-1027 enediyne antitumor antibiotic (Proc. Natl. Acad. Sci. U.S.A. 2008, 105, 494-499)

27. Discovery of the enediyne polyketide synthases as a new class of phosphopantetheinylating iterative polyketide synthases from the biosynthesis of the C-1027, neocarzinostatin, and maduropeptin enediyne antitumor antibiotics (Proc. Natl. Acad. Sci. U.S.A. 2008, 105, 1460-1465)

28. Characterization of SgcC as a FAD-dependent monooxygenase that acts on a carrier protein-tethered substrate for the biosynthesis of the C-1027 enediyne antitumor antibiotic (J. Am. Chem. Soc. 2008, 130, 6616-6623)

29. Characterization of NcsB1 as a promiscuous O-methyltransferase for the biosynthesis of the neocarzinostatin enediyne antitumor antibiotic (J. Biol. Chem. 2008, 283, 14694-14702)

30. Determination of the structure of the NcsB1 O-methyltransferase for the biosynthesis of the neocarzinostatin enediyne antitumor antibiotic (J. Biol. Chem. 2008, 283, 14694-14702)

31. Discovery of the role of tandem acyl carrier protein domains in polyunsaturated fatty acid biosynthesis (J. Am. Chem. Soc. 2008, 130, 6336-6337)

32. Discovery of a free-standing condensation enzyme that can catalyze both C-O ester and C-N amide bond formation for the biosynthesis of the C-1027 enediyne antitumor antibiotic (Prod. natl. Acad. Sci. 2009, 106, 4183-4188)

33. Characterization of TlmK as a glycosyltransferase for tallysomycin biosynthesis (J. Biol. Chem. 2009, 284, 8256-8264)

34. Characterization of TtmM as a a-ketoglutarate-dependent hydroxylase for tautomycin biosynthesis (Org. Lett. 2009, 11, 1639-1642)

35. Discovery of LnmK, a bifunctional acyltransferase/decarboxylase, as the missing link for b-alkylation in polyketide biosynthesis (J. Am.  Chem. Soc. 2009, 132, 6900-6901)

36. Discovery of FdmM and FdmM1 as cofactor-independent novel oxygenases for fredericamycin biosynthesis (J. Biol. Chem. 2009, 284, 24735-24743)

37. Characterization of SgcE6 as a FAD reductase component for the FAD-dependent halogenations and hydroxylation steps in the biosynthesis of the C-1027 enediyne antitumor antibiotic (FEMS Microbiol. Lett. 2009, 300, 237-241)

38. Characterization of SgcF as an epoxide hydrolase to yield an (R)-vicinal diol providing direct evidence for its intermediacy in enediyne core biosynthesis (J. Am. Chem. Soc. 2009, 131, 16410-16417)

39. Characterization of TlmH as a a-ketoglutarate-dependent hydroxylase for tallysomycin biosynthesis (Mol. BioSyst. 2010, 6, 349-256)

40. Characterization of TtnDF as putative decarboxylase and dehydratase for tautomycetin biosynthesis (J. Am. Chem. Soc. 2010, 132, 6663-6671)

41. Establishment of the common PKSE chemistry for the biosynthesis of both 9- and 10-membered enediyne cores with the PKSE accessory enzymes directing enediyne core pathway divergence (Proc. Natl. Acad. Sci. U.S.A. 2010, 107, 11331-11335)

42. Discovery of the MdpB1 C-methyltransferase that acts on a CoA-tethered aromatic substrate for biosynthesis of the maduropeptin enediyne antitumor antibiotic (J. Am. Chem. Soc. 2010, 132, 12534-12536)

43. Discovery of NcsF2 and SgcF as a pair of complementary epoxide hydrolases and demonstration of their utilities in preparation of an optically pure diol from a racemic epoxide substrate (Org. Lett. 2010, 12, 3816-3819)

44. Characterization of FdmV as an amide synthase for fredericamycin biosynthesis (J. Biol. Chem. 2010, 285, 38853-38860)

45. Discovery of LnmKLM to install the b-branch for leinamycin biosynthesis (Org. Lett. 2011, 13, 498-501)