Biosynthesis of sterols and ecdysteroids in Ajuga hairy roots
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, 3alpha-, 4alpha-, and 4beta-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 5beta-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 5alpha,6alpha-epoxide. Moreover, 7-dehydrocholesterol, 3beta-hydroxy-5beta-cholest-7-en-6-one (5beta-ketol), and 3beta,14alpha-dihydroxy-5beta-cholest-7-en-6-one (5beta-ketodiol) were converted into 20-hydroxyecdysone. Thus, the pathway cholesterol-->7-dehydrocholesterol-->7-dehydrocholesterol 5alpha,6alpha-epoxide-->5beta-ketol-->5beta-k etodiol is proposed for the early stages of 20-hydroxyecdysone biosynthesis. 3beta-Hydroxy-5beta-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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To date, only the hairy roots of Ajuga reptans var. atropurpurea consistently and reliably produce ecdysteroids.
“To date, only the hairy roots of Ajuga reptans var.atropurpurea [167] consistently and reliably produce ecdysteroids, but the culture is too expensive to provide a commercial source of 20E.”
From Ecdysteroids (2021) · cites this paper as [167]
Phytoecdysteroids originate from the precursor cholesterol.
“phytoecdysteroids originate from the precursor cholesterol,49”
Cites
Comprehensive insect physiology, biochemistry and pharmacology
Hoilmann, D. (1985) · International Journal of Biochemistry · doi:10.1016/0020-711x(85)90023-0 · Cited by 1,401 · see citing works
Ecological Chemistry and Biochemistry of Plant Terpenoids.
Meurer‐Grimes, B., Harborne, J. B., & Tomás-Barberán, F. A. (1992) · Brittonia · doi:10.2307/2806846 · Cited by 337 · see citing works
Lund, E. G., Kerr, T. A., Sakai, J., Li, W.-P., & Russell, D. W. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.51.34316 · Cited by 335 · see citing works
Ecological chemistry and biochemistry of plant terpenoids
Harborne, J. B., & Tomás-Barberán, F. A. (1991) · Cited by 251 · see citing works
Recent advances in our knowledge of ecdysteroid biosynthesis in insects and crustaceans
Grieneisen, M. L. (1994) · Insect Biochemistry and Molecular Biology · doi:10.1016/0965-1748(94)90078-7 · Cited by 184 · see citing works
Star, A. E., Reinhold, L. A., & Liwschitz, Y. (1973) · Bulletin of the Torrey Botanical Club · doi:10.2307/2484808 · Cited by 172 · see citing works
Feyereisen, R., & Durst, F. (1978) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1978.tb12420.x · Cited by 135 · see citing works
Insect allelochemicals from Ajuga plants
Camps, F., & Coll, J. (1993) · Phytochemistry · doi:10.1016/0031-9422(93)85139-i · Cited by 103 · see citing works
Smith, S. L., Bollenbacher, W. E., Cooper, D. Y., Schleyer, H., Wielgus, J. J., & Gilbert, L. I. (1979) · Molecular and Cellular Endocrinology · doi:10.1016/0303-7207(79)90033-9 · Cited by 98 · see citing works
The Biosynthesis of Plant Sterols
Goad, L. J. (1977) · Springer eBooks · doi:10.1007/978-3-642-66632-2_8 · Cited by 92 · see citing works
Studies of Cholesterol Biosynthesis
Steinberg, D. H., & Avigan, J. (1960) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(20)81322-2 · Cited by 84 · see citing works
Biosynthesis and distribution of insect-molting hormones in plants—A review
Adler, J. H., & Grebenok, R. J. (1995) · Lipids · doi:10.1007/bf02537830 · Cited by 79 · see citing works
Kappler, C., Kabbouh, M., Hétru, C., Durst, F., & Hoffmann, J. A. (1988) · Journal of Steroid Biochemistry · doi:10.1016/0022-4731(88)90330-5 · Cited by 72 · see citing works
Production of phytoecdysteroids by hairy root cultures of Ajuga reptans var. atropurpurea.
Matsumoto, T., & Tanaka, N. (1991) · Agricultural and Biological Chemistry · doi:10.1271/bbb1961.55.1019 · Cited by 72 · see citing works
Svoboda, J. A., & Robbins, W. E. (1971) · Lipids · doi:10.1007/bf02531326 · Cited by 52 · see citing works
Ecdysteroid biosynthesis during the ontogeny of spinach leaves
Grebenok, R. J., & Adler, J. H. (1993) · Phytochemistry · doi:10.1016/0031-9422(93)85514-r · Cited by 48 · see citing works
Studies on the C-2 hydroxylation of 2-deoxyecdysone in Locusta migratoria
Kappler, C., Kabbouh, M., Durst, F., et al. (1986) · Insect Biochemistry · doi:10.1016/0020-1790(86)90074-0 · Cited by 45 · see citing works
Ecdysone biosynthesis: Pathways, enzymes, and the early steps problem
Warren, J. T., & Hétru, C. (1990) · Invertebrate Reproduction & Development · doi:10.1080/07924259.1990.9672131 · Cited by 42 · see citing works
Structure of cyasterone, a novel C29 insect-moulting substance from
TAKEMOTO, T., Hikino, Y., Nomoto, K., & Hikino, H. (1967) · Tetrahedron Letters · doi:10.1016/s0040-4039(01)89896-8 · Cited by 40 · see citing works
Mechanism of formation of the A/B cis ring junction of ecdysteroids in Polypodium vulgare
Davies, T. G. E., Lockley, W. J. S., Boid, R., Rees, H. H., & Goodwin, T. W. (1980) · Biochemical Journal · doi:10.1042/bj1900537 · Cited by 38 · see citing works
Grieneisen, M. L., Warren, J. T., Sakurai, S., & Gilbert, L. I. (1991) · Insect Biochemistry · doi:10.1016/0020-1790(91)90062-j · Cited by 38 · see citing works
Formation of the A/B cis ring junction of ecdysteroids in the locust, Schistocerca gregaria
Davies, T. G. E., Dinan, L. N., Lockley, W. J., Rees, H. H., & Goodwin, T. W. (1981) · Biochemical Journal · doi:10.1042/bj1940053 · Cited by 37 · see citing works
Bollenbacher, W. E., Galbraith, M. N., Gilbert, L. I., & Horn, D. (1977) · Steroids · doi:10.1016/0039-128x(77)90108-8 · Cited by 37 · see citing works
Stereoselective synthesis and solvolytic behavior of the isomeric 7-dehydrocholesterol 5,6-oxides
Michaud, D. P., Nashed, N. T., & Jerina, D. M. (1985) · The Journal of Organic Chemistry · doi:10.1021/jo00211a009 · Cited by 34 · see citing works
Structure of sengosterone, a novel C29 insect-moulting substance from
Hikino, H., Nomoto, K., & Takemoto, T. (1969) · Tetrahedron Letters · doi:10.1016/s0040-4039(01)87902-8 · Cited by 33 · see citing works
Nagakari, M., Kushiro, T., Matsumoto, T., Tanaka, N., Kakinuma, S., & Fujimoto, Y. (1994) · Phytochemistry · doi:10.1016/s0031-9422(00)90461-8 · Cited by 32 · see citing works
Self-citationSynthesis of a labelled putative precursor of ecdysone—II
Haag, T., Meister, M. F., Hétru, C., Kappler, C., Nakatani, Y., Beaucourt, J. P., Rousseau, B., & Luu, B. (1987) · Insect Biochemistry · doi:10.1016/0020-1790(87)90071-0 · Cited by 31 · see citing works
Structure of capitasterone, a novel C29 insect-moulting substance from
Takemoto, T., Nomoto, K., Hikino, Y., & Hikino, H. (1968) · Tetrahedron Letters · doi:10.1016/s0040-4039(00)72794-8 · Cited by 30 · see citing works
Ecdysone 20-monooxygenase, a cytochrome P450 enzyme from spinach, Spinacia oleracea
Grebenok, R. J., Galbraith, D. W., Benveniste, I., & Feyereisen, R. (1996) · Phytochemistry · doi:10.1016/0031-9422(96)00094-5 · Cited by 29 · see citing works
SOUZA, N. J. D., Ghisalberti, E. L., Rees, H. H., & Goodwin, T. W. (1970) · Phytochemistry · doi:10.1016/s0031-9422(00)85315-7 · Cited by 29 · see citing works
Biosynthesis of ecdysteroids in Zea mays
Devarenne, T. P., Sen-Michael, B., & Adler, J. H. (1995) · Phytochemistry · doi:10.1016/0031-9422(95)00435-a · Cited by 28 · see citing works
Biosynthesis of 24β-alkyl-Δ25-sterols in hairy roots of Ajuga reptans var. atropurpurea
Yagi, T., Morisaki, M., Kushiro, T., Yoshida, H., & Fujimoto, Y. (1996) · Phytochemistry · doi:10.1016/0031-9422(95)00766-0 · Cited by 27 · see citing works
Self-citationNeo-clerodane diterpenes from Ajuga ciliata var. villosior.
Shimomura, H., Sashida, Y., & Ogawa, K. (1989) · Chemical and Pharmaceutical Bulletin · doi:10.1248/cpb.37.988 · Cited by 27 · see citing works
Stereospecificity of sterol biosynthesis in Calendula officinalis flowers
Sliwowski, J. K., & Kasprzyk, Z. (1974) · Phytochemistry · doi:10.1016/0031-9422(74)80306-7 · Cited by 26 · see citing works
Goad, L. J., Gibbons, G. F., Bolger, L. M., et al. (1969) · Biochemical Journal · doi:10.1042/bj1140885 · Cited by 25 · see citing works
Faux, A.-M., Galbraith, M. N., Horn, D. H. S., Middleton, E. J., & Thomson, J. A. (1970) · Journal of the Chemical Society D Chemical Communications · doi:10.1039/c29700000243 · Cited by 25 · see citing works
7,8-dihydroajugasterone C, norhygrine and other constituents of Nierembergia hippomanica
Pomilio, A. B., González, M. D., & Eceizabarrena, C. C. (1996) · Phytochemistry · doi:10.1016/0031-9422(95)00731-8 · Cited by 23 · see citing works
Biosynthesis of ecdysterone from cholesterol in Taxus baccata
Lloyd-Jones, J. G., Rees, H. H., & Goodwin, T. W. (1973) · Phytochemistry · doi:10.1016/s0031-9422(00)84445-3 · Cited by 23 · see citing works
Greenwood, D. R., Dinan, L. N., & Rees, H. H. (1984) · Biochemical Journal · doi:10.1042/bj2170783 · Cited by 22 · see citing works
Campagnoni, C. W., Holmlund, C. E., & Whittaker, N. F. (1977) · Archives of Biochemistry and Biophysics · doi:10.1016/0003-9861(77)90465-9 · Cited by 22 · see citing works
Inhibition of Sterol Biosynthesis in Chlorella sorokiniana by Triparanol
Chan, J. T., Patterson, G. W., Dutky, S. R., et al. (1974) · PLANT PHYSIOLOGY · doi:10.1104/pp.53.2.244 · Cited by 22 · see citing works
Nagakari, M., Kushiro, T., Yagi, T., Tanaka, N., Matsumoto, T., Kakinuma, S., & Fujimoto, Y. (1994) · Journal of the Chemical Society Chemical Communications · doi:10.1039/c39940001761 · Cited by 21 · see citing works
Self-citationFujimoto, Y., Miyasaka, S., Ikeda, T., et al. (1985) · Journal of the Chemical Society Chemical Communications · doi:10.1039/c39850000010 · Cited by 20 · see citing works
Self-citationHyodo, R., & Fujimoto, Y. (2000) · Phytochemistry · doi:10.1016/s0031-9422(00)00018-2 · Cited by 19 · see citing works
Self-citationGalli-Kienle, M., Varma, R. K., Mulheirn, L. J., et al. (1973) · Journal of the American Chemical Society · doi:10.1021/ja00787a046 · Cited by 19 · see citing works
Further characterization of the 2-deoxyecdysone C-2 hydroxylase from Locusta migratoria
Kabbouh, M., Kappler, C., Hétru, C., et al. (1987) · Insect Biochemistry · doi:10.1016/0020-1790(87)90088-6 · Cited by 19 · see citing works
Fujimoto, Y., Kushiro, T., & Nakamura, K. (1997) · Tetrahedron Letters · doi:10.1016/s0040-4039(97)00432-2 · Cited by 16 · see citing works
Self-citationヒキノ, ヒ., 野本, 享., & 竹本, 常. (1971) · Chemical and Pharmaceutical Bulletin · Cited by 14 · see citing works
Biosynthesis of 20-hydroxyecdysone in Ajuga hairy roots
Ohyama, K., Kushiro, T., Nakamura, K., & Fujimoto, Y. (1999) · Bioorganic & Medicinal Chemistry · doi:10.1016/s0968-0896(99)00243-6 · Cited by 13 · see citing works
Self-citationBiosynthesis of Ecdysterone from cholesterol by a plant
Heftmanń, E., Sauer, H. H., & Bennett, R. D. (1968) · Die Naturwissenschaften · doi:10.1007/bf00593412 · Cited by 13 · see citing works
Joly, R. A., Svahn, C. M., Bennett, R. D., & Heftmanń, E. (1969) · Phytochemistry · doi:10.1016/s0031-9422(00)88076-0 · Cited by 13 · see citing works
Biosynthesis of 20-hydroxyecdysone in Ajuga hairy roots: Stereochemistry of C-25 hydroxylation
Nakagawa, T., Hara, N., & Fujimoto, Y. (1997) · Tetrahedron Letters · doi:10.1016/s0040-4039(97)00433-4 · Cited by 12 · see citing works
Self-citationFujimoto, Y., Hiramoto, M., Kakinuma, S., et al. (1989) · Steroids · doi:10.1016/0039-128x(89)90026-3 · Cited by 12 · see citing works
Self-citationTen years of ecdysone workshops
Hoffmann, J. A. (1986) · Insect Biochemistry · doi:10.1016/0020-1790(86)90071-5 · Cited by 11 · see citing works
Boid, R., Rees, H. H., & Goodwin, T. W. (1975) · Biochemie und Physiologie der Pflanzen · doi:10.1016/s0015-3796(17)30100-2 · Cited by 10 · see citing works
Tomita, Y., & Sakurai, E. (1974) · Journal of the Chemical Society Chemical Communications · doi:10.1039/c39740000434 · Cited by 9 · see citing works
Biosynthesis of the Insect-Moulting Hormone Cyasterone in the Plant Cyathula capitata
Boid, R., Rees, H. H., & Goodwin, T. W. (1974) · Biochemical Society Transactions · doi:10.1042/bst0021066 · Cited by 9 · see citing works
Biosynthesis of ponasterone A and ecdysterone from cholesterol in Podocarpus macrophyllus
Hikino, H., KOHAMA, T., & Takemoto, T. (1970) · Phytochemistry · doi:10.1016/s0031-9422(00)85149-3 · Cited by 8 · see citing works
Fujimoto, Y., Nagakari, M., Ikuina, Y., Ikekawa, N., & Kakinuma, S. (1991) · Journal of the Chemical Society Chemical Communications · doi:10.1039/c39910000688 · Cited by 7 · see citing works
Self-citationGreenwood, D. R., & Rees, H. H. (1982) · Biochemical Journal · doi:10.1042/bj2080857 · Cited by 7 · see citing works
Yagi, T., Kobayashi, N., Morisaki, M., et al. (1994) · Chemical and Pharmaceutical Bulletin · doi:10.1248/cpb.42.680 · Cited by 7 · see citing works
Self-citationFaux, A. F., Horn, D. H. S., Kinnear, J. F., et al. (1979) · Insect Biochemistry · doi:10.1016/0020-1790(79)90033-7 · Cited by 6 · see citing works
Mechanism of C-2 Hydroxylation during the Biosynthesis of 20-Hydroxyecdysone in Ajuga Hairy Roots.
Nomura, K., & Fujimoto, Y. (2000) · Chemical and Pharmaceutical Bulletin · doi:10.1248/cpb.48.344 · Cited by 5 · see citing works
Self-citationStereochemistry of Hydrogen Introduction at C-25 during Cholesterol Biosynthesis in Higher Plants.
NAGANO, M., Yamada, J., YAZAKI-YAGI, T., et al. (1997) · Chemical and Pharmaceutical Bulletin · doi:10.1248/cpb.45.944 · Cited by 4 · see citing works
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Gilbert, L. I., Rybczynski, R., & Warren, J. T. (2002) · Annual Review of Entomology · doi:10.1146/annurev.ento.47.091201.145302 · Cited by 532 · see citing works
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Dinan, L. (2001) · Phytochemistry · doi:10.1016/s0031-9422(01)00078-4 · Cited by 392 · see citing works
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Bakrim, A., Maria, A., Sayah, F., Lafont, R., & Takvorian, N. (2008) · Plant Physiology and Biochemistry · doi:10.1016/j.plaphy.2008.06.002 · Cited by 63 · see citing works
Lafont, R., Balducci, C., & Dinan, L. (2021) · Encyclopedia · doi:10.3390/encyclopedia1040096 · Cited by 49 · see citing works
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- Authors
- Fujimoto, Y., Ohyama, K., Nomura, K., Hyodo, R., Takahashi, K., Yamada, J., & Morisaki, M.
- Affiliations
- Tokyo Institute of Technology (JP)
- Kyoritsu College of Pharmacy Shibakoen Tokyo 105‐8512 Japan
- Corresponding author
- Fujimoto, Yoshinori
- Venue
- Lipids
- Publisher
- Wiley
- Published
- 2000-03
- Volume
- 35
- Issue
- 3
- Pages
- 279-288
- ISSN
- 0024-4201 (ISSN-L)
- 1558-9307
- OpenAlex
- W2029708628
- Cited by
- 45
- References
- 64
- Citations by year
- 2026: 2 · 2025: 2 · 2024: 2 · 2023: 3 · 2022: 3 · 2021: 3 · 2019: 1 · 2018: 1 · 2017: 3 · 2016: 2 · 2015: 2 · 2013: 1
- FWCI
- 0.84
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Lipids (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant biochemistry and biosynthesis
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Steroid Chemistry and Biochemistry
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Plant tissue culture and regeneration
- Keywords
- sterol biosynthesis, ecdysteroid biosynthesis, hairy roots, 20-hydroxyecdysone, clerosterol, cholesterol, 7-dehydrocholesterol
- MeSH terms
- Carbon Isotopes, Deuterium, Models, Chemical, Plants, Medicinal, Stereoisomerism, Steroids, Sterols, Stigmasterol, Plant Roots, Ecdysteroids
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