Oxidation and cyclization of Casbene in the biosynthesis of Euphorbia factors from mature seeds of Euphorbia lathyris L

Dan Luo; Roberta Callari; Britta Hamberger; Sileshi Gizachew Wubshet; Morten Thrane Nielsen; Johan Andersen-Ranberg; Björn M Hallström; Federico Cozzi; Harald Heider; Birger Lindberg Møller; Dan Stærk; Björn Robert Hamberger

doi:10.1073/pnas.1607504113

Oxidation and cyclization of Casbene in the biosynthesis of Euphorbia factors from mature seeds of Euphorbia lathyris L

Dan Luo, Roberta Callari, Britta Hamberger, Sileshi Gizachew Wubshet, Morten Thrane Nielsen, Johan Andersen-Ranberg, Björn M Hallström, Federico Cozzi, Harald Heider, Birger Lindberg Møller, Dan Stærk, Björn Robert Hamberger

42 Citationer (Scopus)

Abstract

The seed oil of Euphorbia lathyris L. contains a series of macrocyclic diterpenoids known as Euphorbia factors. They are the current industrial source of ingenol mebutate, which is approved for the treatment of actinic keratosis, a precancerous skin condition. Here, we report an alcohol dehydrogenase-mediated cyclization step in the biosynthetic pathway of Euphorbia factors, illustrating the origin of the intramolecular carbon-carbon bonds present in lathyrane and ingenane diterpenoids. This unconventional cyclization describes the ring closure of the macrocyclic diterpene casbene. Through transcriptomic analysis of E. lathyris L. mature seeds and in planta functional characterization, we identified three enzymes involved in the cyclization route from casbene to jolkinol C, a lathyrane diterpene. These enzymes include two cytochromes P450 from the CYP71 clan and an alcohol dehydrogenase (ADH). CYP71D445 and CYP726A27 catalyze regio-specific 9-oxidation and 5-oxidation of casbene, respectively. When coupled with these P450-catalyzed monooxygenations, E. lathyris ADH1 catalyzes dehydrogenation of the hydroxyl groups, leading to the subsequent rearrangement and cyclization. The discovery of this nonconventional cyclization may provide the key link to complete elucidation of the biosynthetic pathways of ingenol mebutate and other bioactive macrocyclic diterpenoids.

Originalsprog	Engelsk
Tidsskrift	Proceedings of the National Academy of Sciences of the United States of America
Vol/bind	113
Udgave nummer	34
Sider (fra-til)	E5082-E5089
Antal sider	8
ISSN	0027-8424
DOI	https://doi.org/10.1073/pnas.1607504113
Status	Udgivet - 23 aug. 2016

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10.1073/pnas.1607504113

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Luo, D., Callari, R., Hamberger, B., Wubshet, S. G., Nielsen, M. T., Andersen-Ranberg, J., Hallström, B. M., Cozzi, F., Heider, H., Møller, B. L., Stærk, D., & Hamberger, B. R. (2016). Oxidation and cyclization of Casbene in the biosynthesis of Euphorbia factors from mature seeds of Euphorbia lathyris L. Proceedings of the National Academy of Sciences of the United States of America, 113(34), E5082-E5089. https://doi.org/10.1073/pnas.1607504113

Oxidation and cyclization of Casbene in the biosynthesis of Euphorbia factors from mature seeds of Euphorbia lathyris L. / Luo, Dan; Callari, Roberta; Hamberger, Britta et al.

I: Proceedings of the National Academy of Sciences of the United States of America, Bind 113, Nr. 34, 23.08.2016, s. E5082-E5089.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

Luo, D, Callari, R, Hamberger, B, Wubshet, SG, Nielsen, MT, Andersen-Ranberg, J, Hallström, BM, Cozzi, F, Heider, H, Møller, BL , Stærk, D & Hamberger, BR 2016, 'Oxidation and cyclization of Casbene in the biosynthesis of Euphorbia factors from mature seeds of Euphorbia lathyris L', Proceedings of the National Academy of Sciences of the United States of America, bind 113, nr. 34, s. E5082-E5089. https://doi.org/10.1073/pnas.1607504113

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title = "Oxidation and cyclization of Casbene in the biosynthesis of Euphorbia factors from mature seeds of Euphorbia lathyris L",

abstract = "The seed oil of Euphorbia lathyris L. contains a series of macrocyclic diterpenoids known as Euphorbia factors. They are the current industrial source of ingenol mebutate, which is approved for the treatment of actinic keratosis, a precancerous skin condition. Here, we report an alcohol dehydrogenase-mediated cyclization step in the biosynthetic pathway of Euphorbia factors, illustrating the origin of the intramolecular carbon-carbon bonds present in lathyrane and ingenane diterpenoids. This unconventional cyclization describes the ring closure of the macrocyclic diterpene casbene. Through transcriptomic analysis of E. lathyris L. mature seeds and in planta functional characterization, we identified three enzymes involved in the cyclization route from casbene to jolkinol C, a lathyrane diterpene. These enzymes include two cytochromes P450 from the CYP71 clan and an alcohol dehydrogenase (ADH). CYP71D445 and CYP726A27 catalyze regio-specific 9-oxidation and 5-oxidation of casbene, respectively. When coupled with these P450-catalyzed monooxygenations, E. lathyris ADH1 catalyzes dehydrogenation of the hydroxyl groups, leading to the subsequent rearrangement and cyclization. The discovery of this nonconventional cyclization may provide the key link to complete elucidation of the biosynthetic pathways of ingenol mebutate and other bioactive macrocyclic diterpenoids.",

author = "Dan Luo and Roberta Callari and Britta Hamberger and Wubshet, {Sileshi Gizachew} and Nielsen, {Morten Thrane} and Johan Andersen-Ranberg and Hallstr{\"o}m, {Bj{\"o}rn M} and Federico Cozzi and Harald Heider and M{\o}ller, {Birger Lindberg} and Dan St{\ae}rk and Hamberger, {Bj{\"o}rn Robert}",

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AU - Luo, Dan

AU - Callari, Roberta

AU - Hamberger, Britta

AU - Wubshet, Sileshi Gizachew

AU - Nielsen, Morten Thrane

AU - Andersen-Ranberg, Johan

AU - Hallström, Björn M

AU - Cozzi, Federico

AU - Heider, Harald

AU - Møller, Birger Lindberg

AU - Stærk, Dan

AU - Hamberger, Björn Robert

PY - 2016/8/23

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N2 - The seed oil of Euphorbia lathyris L. contains a series of macrocyclic diterpenoids known as Euphorbia factors. They are the current industrial source of ingenol mebutate, which is approved for the treatment of actinic keratosis, a precancerous skin condition. Here, we report an alcohol dehydrogenase-mediated cyclization step in the biosynthetic pathway of Euphorbia factors, illustrating the origin of the intramolecular carbon-carbon bonds present in lathyrane and ingenane diterpenoids. This unconventional cyclization describes the ring closure of the macrocyclic diterpene casbene. Through transcriptomic analysis of E. lathyris L. mature seeds and in planta functional characterization, we identified three enzymes involved in the cyclization route from casbene to jolkinol C, a lathyrane diterpene. These enzymes include two cytochromes P450 from the CYP71 clan and an alcohol dehydrogenase (ADH). CYP71D445 and CYP726A27 catalyze regio-specific 9-oxidation and 5-oxidation of casbene, respectively. When coupled with these P450-catalyzed monooxygenations, E. lathyris ADH1 catalyzes dehydrogenation of the hydroxyl groups, leading to the subsequent rearrangement and cyclization. The discovery of this nonconventional cyclization may provide the key link to complete elucidation of the biosynthetic pathways of ingenol mebutate and other bioactive macrocyclic diterpenoids.

AB - The seed oil of Euphorbia lathyris L. contains a series of macrocyclic diterpenoids known as Euphorbia factors. They are the current industrial source of ingenol mebutate, which is approved for the treatment of actinic keratosis, a precancerous skin condition. Here, we report an alcohol dehydrogenase-mediated cyclization step in the biosynthetic pathway of Euphorbia factors, illustrating the origin of the intramolecular carbon-carbon bonds present in lathyrane and ingenane diterpenoids. This unconventional cyclization describes the ring closure of the macrocyclic diterpene casbene. Through transcriptomic analysis of E. lathyris L. mature seeds and in planta functional characterization, we identified three enzymes involved in the cyclization route from casbene to jolkinol C, a lathyrane diterpene. These enzymes include two cytochromes P450 from the CYP71 clan and an alcohol dehydrogenase (ADH). CYP71D445 and CYP726A27 catalyze regio-specific 9-oxidation and 5-oxidation of casbene, respectively. When coupled with these P450-catalyzed monooxygenations, E. lathyris ADH1 catalyzes dehydrogenation of the hydroxyl groups, leading to the subsequent rearrangement and cyclization. The discovery of this nonconventional cyclization may provide the key link to complete elucidation of the biosynthetic pathways of ingenol mebutate and other bioactive macrocyclic diterpenoids.

U2 - 10.1073/pnas.1607504113

DO - 10.1073/pnas.1607504113

M3 - Journal article

C2 - 27506796

SN - 0027-8424

VL - 113

SP - E5082-E5089

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 34

ER -

Oxidation and cyclization of Casbene in the biosynthesis of Euphorbia factors from mature seeds of Euphorbia lathyris L

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