Biosynthesis of sterols and ecdysteroids in Ajuga hairy roots

Yoshinori Fujimoto, Kiyoshi Ohyama, Keiko Nomura, Ryo Hyodo, Kyoko Takahashi, Junko Yamada, Masuo Morisaki

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Hairy roots of Ajuga reptans var. atropurpurea produce clerosterol, 22- dehydroclerosterol, and cholesterol as sterol constituents, and 20- hydroxyecdysone, cyasterone, isocyasterone, and 29-norcyasterone as ecdysteroid constituents. To better understand the biosynthesis of these steroidal compounds, we carried out feeding studies of variously 2H- and 13C-labeled sterol substrates with Ajuga hairy roots. In this article, we review our studies in this field. Feeding of labeled desmosterols, 24- methylenecholesterol, and 13C2-acetate established the mechanism of the biosynthesis of the two C29-sterols and a newly accumulated codisterol, including the metabolic correlation of C-26 and C-27 methyl groups. In Ajuga hairy roots, 3α-, 4α-, and 4β-hydrogens of cholesterol were all retained at their original positions after conversion into 20-hydroxyecdysone, in contrast to the observations in a fern and an insect. Furthermore, the origin of 5β-H of 20-hydroxyecdysone was found to be C-6 hydrogen of cholesterol exclusively, which is inconsistent with the results in the fern and the insect. These data strongly support the intermediacy of 7-dehydrocholesterol 5α,6α-epoxide. Moreover, 7-dehydrocholesterol, 3β-hydroxy-5β-cholest-7- en-6-one (5β-ketol), and 3β,14α-dihydroxy-5β-cholest-7-en-6-one (5β- ketodiol) were converted into 20-hydroxyecdysone. Thus, the pathway cholesterol → 7-dehydrocholesterol → 7-dehydrocholesterol 5α, 6α-epoxide → 5β-ketol → 5β-ketodiol is proposed for the early stages of 20- hydroxyecdysone biosynthesis. 3β-Hydroxy-5β-cholestan-6-one was also incorporated into 20-hydroxyecdysone, suggesting that the introduction of a 7-ene function is not necessarily next to cholesterol. C-25 Hydroxylation during 20-hydroxyecdysone biosynthesis was found to proceed with ca. 70% retention and 30% inversion. Finally, clerosterol was shown to be a precursor of cyasterone and isocyasterone.

Original languageEnglish
Pages (from-to)279-288
Number of pages10
JournalLipids
Volume35
Issue number3
Publication statusPublished - 2000

Fingerprint

Ajuga
Ecdysterone
Ecdysteroids
ecdysterone
ecdysteroids
Biosynthesis
Sterols
sterols
7-dehydrocholesterol
biosynthesis
Cholesterol
cholesterol
Ferns
epoxides
ferns and fern allies
hydrogen
Insects
Hydrogen
Ajuga reptans
Desmosterol

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Food Science
  • Biochemistry

Cite this

Fujimoto, Y., Ohyama, K., Nomura, K., Hyodo, R., Takahashi, K., Yamada, J., & Morisaki, M. (2000). Biosynthesis of sterols and ecdysteroids in Ajuga hairy roots. Lipids, 35(3), 279-288.

Biosynthesis of sterols and ecdysteroids in Ajuga hairy roots. / Fujimoto, Yoshinori; Ohyama, Kiyoshi; Nomura, Keiko; Hyodo, Ryo; Takahashi, Kyoko; Yamada, Junko; Morisaki, Masuo.

In: Lipids, Vol. 35, No. 3, 2000, p. 279-288.

Research output: Contribution to journalArticle

Fujimoto, Y, Ohyama, K, Nomura, K, Hyodo, R, Takahashi, K, Yamada, J & Morisaki, M 2000, 'Biosynthesis of sterols and ecdysteroids in Ajuga hairy roots', Lipids, vol. 35, no. 3, pp. 279-288.
Fujimoto Y, Ohyama K, Nomura K, Hyodo R, Takahashi K, Yamada J et al. Biosynthesis of sterols and ecdysteroids in Ajuga hairy roots. Lipids. 2000;35(3):279-288.
Fujimoto, Yoshinori ; Ohyama, Kiyoshi ; Nomura, Keiko ; Hyodo, Ryo ; Takahashi, Kyoko ; Yamada, Junko ; Morisaki, Masuo. / Biosynthesis of sterols and ecdysteroids in Ajuga hairy roots. In: Lipids. 2000 ; Vol. 35, No. 3. pp. 279-288.
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AU - Fujimoto, Yoshinori

AU - Ohyama, Kiyoshi

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AU - Takahashi, Kyoko

AU - Yamada, Junko

AU - Morisaki, Masuo

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N2 - Hairy roots of Ajuga reptans var. atropurpurea produce clerosterol, 22- dehydroclerosterol, and cholesterol as sterol constituents, and 20- hydroxyecdysone, cyasterone, isocyasterone, and 29-norcyasterone as ecdysteroid constituents. To better understand the biosynthesis of these steroidal compounds, we carried out feeding studies of variously 2H- and 13C-labeled sterol substrates with Ajuga hairy roots. In this article, we review our studies in this field. Feeding of labeled desmosterols, 24- methylenecholesterol, and 13C2-acetate established the mechanism of the biosynthesis of the two C29-sterols and a newly accumulated codisterol, including the metabolic correlation of C-26 and C-27 methyl groups. In Ajuga hairy roots, 3α-, 4α-, and 4β-hydrogens of cholesterol were all retained at their original positions after conversion into 20-hydroxyecdysone, in contrast to the observations in a fern and an insect. Furthermore, the origin of 5β-H of 20-hydroxyecdysone was found to be C-6 hydrogen of cholesterol exclusively, which is inconsistent with the results in the fern and the insect. These data strongly support the intermediacy of 7-dehydrocholesterol 5α,6α-epoxide. Moreover, 7-dehydrocholesterol, 3β-hydroxy-5β-cholest-7- en-6-one (5β-ketol), and 3β,14α-dihydroxy-5β-cholest-7-en-6-one (5β- ketodiol) were converted into 20-hydroxyecdysone. Thus, the pathway cholesterol → 7-dehydrocholesterol → 7-dehydrocholesterol 5α, 6α-epoxide → 5β-ketol → 5β-ketodiol is proposed for the early stages of 20- hydroxyecdysone biosynthesis. 3β-Hydroxy-5β-cholestan-6-one was also incorporated into 20-hydroxyecdysone, suggesting that the introduction of a 7-ene function is not necessarily next to cholesterol. C-25 Hydroxylation during 20-hydroxyecdysone biosynthesis was found to proceed with ca. 70% retention and 30% inversion. Finally, clerosterol was shown to be a precursor of cyasterone and isocyasterone.

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