Generation of Complexity in Fungal Terpene Biosynthesis: Discovery of a Multifunctional Cytochrome P450 in the Fumagillin Pathway

Overview

Journal J Am Chem Soc

Publisher American Chemical Society

Specialty Chemistry

Date 2014 Feb 27

PMID 24568283

Citations 25

Authors

Hsiao-Ching Lin

Yuta Tsunematsu

Sourabh Dhingra

Wei Xu

Manami Fukutomi

Yit-Heng Chooi

David E Cane

Ana M Calvo

Kenji Watanabe

Yi Tang

Affiliations

Soon will be listed here.

Abstract

Fumagillin (1), a meroterpenoid from Aspergillus fumigatus, is known for its antiangiogenic activity due to binding to human methionine aminopeptidase 2. 1 has a highly oxygenated structure containing a penta-substituted cyclohexane that is generated by oxidative cleavage of the bicyclic sesquiterpene β-trans-bergamotene. The chemical nature, order, and biochemical mechanism of all the oxygenative tailoring reactions has remained enigmatic despite the identification of the biosynthetic gene cluster and the use of targeted-gene deletion experiments. Here, we report the identification and characterization of three oxygenases from the fumagillin biosynthetic pathway, including a multifunctional cytochrome P450 monooxygenase, a hydroxylating nonheme-iron-dependent dioxygenase, and an ABM family monooxygenase for oxidative cleavage of the polyketide moiety. Most significantly, the P450 monooxygenase is shown to catalyze successive hydroxylation, bicyclic ring-opening, and two epoxidations that generate the sesquiterpenoid core skeleton of 1. We also characterized a truncated polyketide synthase with a ketoreductase function that controls the configuration at C-5 of hydroxylated intermediates.

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References

Li S, Tietz D, Rutaganira F, Kells P, Anzai Y, Kato F . Substrate recognition by the multifunctional cytochrome P450 MycG in mycinamicin hydroxylation and epoxidation reactions. J Biol Chem. 2012; 287(45):37880-90. PMC: 3488060. DOI: 10.1074/jbc.M112.410340. View

Matsuda Y, Awakawa T, Wakimoto T, Abe I . Spiro-ring formation is catalyzed by a multifunctional dioxygenase in austinol biosynthesis. J Am Chem Soc. 2013; 135(30):10962-5. DOI: 10.1021/ja405518u. View

BIRCH A, Hussain S . Studies in relation to biosynthesis. 38. A preliminary study of fumagillin. J Chem Soc Perkin 1. 1969; 11:1473-4. DOI: 10.1039/j39690001473. View

Bernier S, Lazarus D, Clark E, Doyle B, Labenski M, Thompson C . A methionine aminopeptidase-2 inhibitor, PPI-2458, for the treatment of rheumatoid arthritis. Proc Natl Acad Sci U S A. 2004; 101(29):10768-73. PMC: 490009. DOI: 10.1073/pnas.0404105101. View

Taramino S, Teske B, Oliaro-Bosso S, Bard M, Balliano G . Divergent interactions involving the oxidosqualene cyclase and the steroid-3-ketoreductase in the sterol biosynthetic pathway of mammals and yeasts. Biochim Biophys Acta. 2010; 1801(11):1232-7. PMC: 2946411. DOI: 10.1016/j.bbalip.2010.07.006. View

Mizutani M, Sato F . Unusual P450 reactions in plant secondary metabolism. Arch Biochem Biophys. 2010; 507(1):194-203. DOI: 10.1016/j.abb.2010.09.026. View

KRUGER E, Figg W . TNP-470: an angiogenesis inhibitor in clinical development for cancer. Expert Opin Investig Drugs. 2000; 9(6):1383-96. DOI: 10.1517/13543784.9.6.1383. View

Podust L, Sherman D . Diversity of P450 enzymes in the biosynthesis of natural products. Nat Prod Rep. 2012; 29(10):1251-66. PMC: 3454455. DOI: 10.1039/c2np20020a. View

Molina J, Tourneur M, Sarfati C, Chevret S, de Gouvello A, Gobert J . Fumagillin treatment of intestinal microsporidiosis. N Engl J Med. 2002; 346(25):1963-9. DOI: 10.1056/NEJMoa012924. View

10.

Sin N, Meng L, Wang M, Wen J, Bornmann W, Crews C . The anti-angiogenic agent fumagillin covalently binds and inhibits the methionine aminopeptidase, MetAP-2. Proc Natl Acad Sci U S A. 1997; 94(12):6099-103. PMC: 21008. DOI: 10.1073/pnas.94.12.6099. View

11.

Cane D, LEVIN R . Application of carbon-13 magnetic resonance to isoprenoid biosynthesis. II. Ovalicin and the use of doubly labeled mevalonate. J Am Chem Soc. 1976; 98(5):1183-8. DOI: 10.1021/ja00421a022. View

12.

Cane D, Buchwald S . Application of deuterium magnetic resonance to biosynthetic studies. 1. Biosynthesis of ovalicin. J Am Chem Soc. 1977; 99(18):6132-4. DOI: 10.1021/ja00460a065. View

13.

Auclair K, Hu Z, Little D, de Montellano P, Groves J . Revisiting the mechanism of P450 enzymes with the radical clocks norcarane and spiro[2,5]octane. J Am Chem Soc. 2002; 124(21):6020-7. DOI: 10.1021/ja025608h. View

14.

Kim D, Min J, Ahn S, Lee H, Choi N, Moon S . 5-Demethoxyfumagillol, a potent angiogenesis inhibitor isolated from Aspergillus fumigatus. Chem Pharm Bull (Tokyo). 2004; 52(4):447-50. DOI: 10.1248/cpb.52.447. View

15.

Barriuso J, Nguyen D, Li J, Roberts J, MacNevin G, Chaytor J . Double oxidation of the cyclic nonaketide dihydromonacolin L to monacolin J by a single cytochrome P450 monooxygenase, LovA. J Am Chem Soc. 2011; 133(21):8078-81. DOI: 10.1021/ja201138v. View

16.

Wen T, Zhou Z, Jia G . Synthesis and Characterization of a Metallabenzyne This work was supported by the Hong Kong Research Grant Council and The Hong Kong University of Science and Technology. We thank Professor C. P. Lau for valuable discussions. . Angew Chem Int Ed Engl. 2001; 40(10):1951-1954. View

17.

Wiemann P, Guo C, M Palmer J, Sekonyela R, Wang C, Keller N . Prototype of an intertwined secondary-metabolite supercluster. Proc Natl Acad Sci U S A. 2013; 110(42):17065-70. PMC: 3801025. DOI: 10.1073/pnas.1313258110. View

18.

Kelley L, Sternberg M . Protein structure prediction on the Web: a case study using the Phyre server. Nat Protoc. 2009; 4(3):363-71. DOI: 10.1038/nprot.2009.2. View

19.

Maiya S, Grundmann A, Li X, Li S, Turner G . Identification of a hybrid PKS/NRPS required for pseurotin A biosynthesis in the human pathogen Aspergillus fumigatus. Chembiochem. 2007; 8(14):1736-43. DOI: 10.1002/cbic.200700202. View

20.

Yamamoto H, Katano N, Ooi A, Inoue K . Secologanin synthase which catalyzes the oxidative cleavage of loganin into secologanin is a cytochrome P450. Phytochemistry. 2000; 53(1):7-12. DOI: 10.1016/s0031-9422(99)00471-9. View