Generation of complexity in fungal terpene biosynthesis: Discovery of a multifunctional cytochrome P450 in the fumagillin pathway

H.C. Lin, Y. Tsunematsu, S. Dhingra, W. Xu, M. Fukutomi, Heng Chooi, D.E. Cane, A.M. Calvo, K. Watanabe, Y. Tang

Research output: Contribution to journalArticle

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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. © 2014 American Chemical Society.
Original languageEnglish
Pages (from-to)4426-4436
JournalJournal of the American Chemical Society
Volume136
Issue number11
DOIs
Publication statusPublished - 2014

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Terpenes
Biosynthesis
Mixed Function Oxygenases
Cytochrome P-450 Enzyme System
Genes
Hydroxylation
Epoxidation
Aspergillus
Cyclohexane
Polyketide Synthases
Polyketides
Dioxygenases
Oxygenases
Rubiaceae
Aspergillus fumigatus
Sesquiterpenes
Biosynthetic Pathways
Gene Deletion
Multigene Family
Iron

Cite this

Lin, H.C. ; Tsunematsu, Y. ; Dhingra, S. ; Xu, W. ; Fukutomi, M. ; Chooi, Heng ; Cane, D.E. ; Calvo, A.M. ; Watanabe, K. ; Tang, Y. / Generation of complexity in fungal terpene biosynthesis: Discovery of a multifunctional cytochrome P450 in the fumagillin pathway. In: Journal of the American Chemical Society. 2014 ; Vol. 136, No. 11. pp. 4426-4436.
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Lin, HC, Tsunematsu, Y, Dhingra, S, Xu, W, Fukutomi, M, Chooi, H, Cane, DE, Calvo, AM, Watanabe, K & Tang, Y 2014, 'Generation of complexity in fungal terpene biosynthesis: Discovery of a multifunctional cytochrome P450 in the fumagillin pathway' Journal of the American Chemical Society, vol. 136, no. 11, pp. 4426-4436. https://doi.org/10.1021/ja500881e

Generation of complexity in fungal terpene biosynthesis: Discovery of a multifunctional cytochrome P450 in the fumagillin pathway. / Lin, H.C.; Tsunematsu, Y.; Dhingra, S.; Xu, W.; Fukutomi, M.; Chooi, Heng; Cane, D.E.; Calvo, A.M.; Watanabe, K.; Tang, Y.

In: Journal of the American Chemical Society, Vol. 136, No. 11, 2014, p. 4426-4436.

Research output: Contribution to journalArticle

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T1 - Generation of complexity in fungal terpene biosynthesis: Discovery of a multifunctional cytochrome P450 in the fumagillin pathway

AU - Lin, H.C.

AU - Tsunematsu, Y.

AU - Dhingra, S.

AU - Xu, W.

AU - Fukutomi, M.

AU - Chooi, Heng

AU - Cane, D.E.

AU - Calvo, A.M.

AU - Watanabe, K.

AU - Tang, Y.

PY - 2014

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N2 - 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. © 2014 American Chemical Society.

AB - 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. © 2014 American Chemical Society.

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JO - Journal of the Amercian Chemical Society

JF - Journal of the Amercian Chemical Society

SN - 0002-7863

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