Enhanced secretion of tropane alkaloids in Nicotiana tabacum hairy roots expressing heterologous hyoscyamine-6β-hydroxylase

Suvi T. Häkkinen (Corresponding Author), E. Moyano, R. M. Cusidó, J. Palazón, M. T. Piñol, Kirsi-Marja Oksman-Caldentey

Research output: Contribution to journalArticleScientificpeer-review

56 Citations (Scopus)

Abstract

Hyoscyamine-6β-hydroxylase (H6H; EC 1.14.11.11) catalyses oxidative reactions in the biosynthetic pathway leading from hyoscyamine to the more pharmaceutically valuable tropane alkaloid scopolamine. The h6h gene encoding H6H from Hyoscyamus niger was introduced, under the control of the CaMV 35S promoter, into non-hyoscyamine-producing Nicotiana tabacum and hyoscyamine-producing Hyoscyamus muticus. The transformation was performed using a binary vector system based on Agrobacterium rhizogenes. Production of scopolamine in hairy roots was clearly correlated with the 35S-h6h transcript expression. The engineered N. tabacum and H. muticus hairy roots were studied for the production of scopolamine and other tropane and nicotine alkaloids after feeding the cultures with exogenous hyoscyamine. N. tabacum hairy roots carrying the 35S-h6h transgene showed a more efficient uptake of hyoscyamine from the culture medium and a higher rate of bioconversion of hyoscyamine to scopolamine than those of H. muticus. In particular, the secretion of scopolamine in N. tabacum hairy roots was remarkably high, up to 85% of the total scopolamine being released to the culture medium. Exogenous hyoscyamine also caused changes in nicotine alkaloid accumulation in N. tabacum hairy roots.
Original languageEnglish
Pages (from-to)2611-2618
Number of pages8
JournalJournal of Experimental Botany
Volume56
Issue number420
DOIs
Publication statusPublished - Oct 2005
MoE publication typeA1 Journal article-refereed

Fingerprint

Hyoscyamine
Tropanes
tropane alkaloids
scopolamine
atropine
Mixed Function Oxygenases
Scopolamine Hydrobromide
Alkaloids
Nicotiana tabacum
Tobacco
secretion
Hyoscyamus muticus
Hyoscyamus
nicotine
hyoscyamine (6S)-dioxygenase
Nicotine
Culture Media
alkaloids
culture media
Hyoscyamus niger

Keywords

  • Hairy roots
  • Hyoscyamine-6beta-hydroxylase
  • Hyoscyamus
  • Nicotiana
  • Secretion
  • Tropane alkaloids

Cite this

@article{d156b764504a4cc7b1e821722b29644d,
title = "Enhanced secretion of tropane alkaloids in Nicotiana tabacum hairy roots expressing heterologous hyoscyamine-6β-hydroxylase",
abstract = "Hyoscyamine-6β-hydroxylase (H6H; EC 1.14.11.11) catalyses oxidative reactions in the biosynthetic pathway leading from hyoscyamine to the more pharmaceutically valuable tropane alkaloid scopolamine. The h6h gene encoding H6H from Hyoscyamus niger was introduced, under the control of the CaMV 35S promoter, into non-hyoscyamine-producing Nicotiana tabacum and hyoscyamine-producing Hyoscyamus muticus. The transformation was performed using a binary vector system based on Agrobacterium rhizogenes. Production of scopolamine in hairy roots was clearly correlated with the 35S-h6h transcript expression. The engineered N. tabacum and H. muticus hairy roots were studied for the production of scopolamine and other tropane and nicotine alkaloids after feeding the cultures with exogenous hyoscyamine. N. tabacum hairy roots carrying the 35S-h6h transgene showed a more efficient uptake of hyoscyamine from the culture medium and a higher rate of bioconversion of hyoscyamine to scopolamine than those of H. muticus. In particular, the secretion of scopolamine in N. tabacum hairy roots was remarkably high, up to 85{\%} of the total scopolamine being released to the culture medium. Exogenous hyoscyamine also caused changes in nicotine alkaloid accumulation in N. tabacum hairy roots.",
keywords = "Hairy roots, Hyoscyamine-6beta-hydroxylase, Hyoscyamus, Nicotiana, Secretion, Tropane alkaloids",
author = "H{\"a}kkinen, {Suvi T.} and E. Moyano and Cusid{\'o}, {R. M.} and J. Palaz{\'o}n and Pi{\~n}ol, {M. T.} and Kirsi-Marja Oksman-Caldentey",
year = "2005",
month = "10",
doi = "10.1093/jxb/eri253",
language = "English",
volume = "56",
pages = "2611--2618",
journal = "Journal of Experimental Botany",
issn = "0022-0957",
publisher = "Oxford University Press",
number = "420",

}

Enhanced secretion of tropane alkaloids in Nicotiana tabacum hairy roots expressing heterologous hyoscyamine-6β-hydroxylase. / Häkkinen, Suvi T. (Corresponding Author); Moyano, E.; Cusidó, R. M.; Palazón, J.; Piñol, M. T.; Oksman-Caldentey, Kirsi-Marja.

In: Journal of Experimental Botany, Vol. 56, No. 420, 10.2005, p. 2611-2618.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Enhanced secretion of tropane alkaloids in Nicotiana tabacum hairy roots expressing heterologous hyoscyamine-6β-hydroxylase

AU - Häkkinen, Suvi T.

AU - Moyano, E.

AU - Cusidó, R. M.

AU - Palazón, J.

AU - Piñol, M. T.

AU - Oksman-Caldentey, Kirsi-Marja

PY - 2005/10

Y1 - 2005/10

N2 - Hyoscyamine-6β-hydroxylase (H6H; EC 1.14.11.11) catalyses oxidative reactions in the biosynthetic pathway leading from hyoscyamine to the more pharmaceutically valuable tropane alkaloid scopolamine. The h6h gene encoding H6H from Hyoscyamus niger was introduced, under the control of the CaMV 35S promoter, into non-hyoscyamine-producing Nicotiana tabacum and hyoscyamine-producing Hyoscyamus muticus. The transformation was performed using a binary vector system based on Agrobacterium rhizogenes. Production of scopolamine in hairy roots was clearly correlated with the 35S-h6h transcript expression. The engineered N. tabacum and H. muticus hairy roots were studied for the production of scopolamine and other tropane and nicotine alkaloids after feeding the cultures with exogenous hyoscyamine. N. tabacum hairy roots carrying the 35S-h6h transgene showed a more efficient uptake of hyoscyamine from the culture medium and a higher rate of bioconversion of hyoscyamine to scopolamine than those of H. muticus. In particular, the secretion of scopolamine in N. tabacum hairy roots was remarkably high, up to 85% of the total scopolamine being released to the culture medium. Exogenous hyoscyamine also caused changes in nicotine alkaloid accumulation in N. tabacum hairy roots.

AB - Hyoscyamine-6β-hydroxylase (H6H; EC 1.14.11.11) catalyses oxidative reactions in the biosynthetic pathway leading from hyoscyamine to the more pharmaceutically valuable tropane alkaloid scopolamine. The h6h gene encoding H6H from Hyoscyamus niger was introduced, under the control of the CaMV 35S promoter, into non-hyoscyamine-producing Nicotiana tabacum and hyoscyamine-producing Hyoscyamus muticus. The transformation was performed using a binary vector system based on Agrobacterium rhizogenes. Production of scopolamine in hairy roots was clearly correlated with the 35S-h6h transcript expression. The engineered N. tabacum and H. muticus hairy roots were studied for the production of scopolamine and other tropane and nicotine alkaloids after feeding the cultures with exogenous hyoscyamine. N. tabacum hairy roots carrying the 35S-h6h transgene showed a more efficient uptake of hyoscyamine from the culture medium and a higher rate of bioconversion of hyoscyamine to scopolamine than those of H. muticus. In particular, the secretion of scopolamine in N. tabacum hairy roots was remarkably high, up to 85% of the total scopolamine being released to the culture medium. Exogenous hyoscyamine also caused changes in nicotine alkaloid accumulation in N. tabacum hairy roots.

KW - Hairy roots

KW - Hyoscyamine-6beta-hydroxylase

KW - Hyoscyamus

KW - Nicotiana

KW - Secretion

KW - Tropane alkaloids

U2 - 10.1093/jxb/eri253

DO - 10.1093/jxb/eri253

M3 - Article

C2 - 16105856

VL - 56

SP - 2611

EP - 2618

JO - Journal of Experimental Botany

JF - Journal of Experimental Botany

SN - 0022-0957

IS - 420

ER -