Alkaloid production in Duboisia hybrid hairy roots and plants overexpressing the h6h gene

Javier Palazon, Elisabet Moyano, Rosa M. Cusido, M. Bonfill, Kirsi-Marja Oksman-Caldentey, M. Teresa Pinol (Corresponding Author)

Research output: Contribution to journalArticleScientificpeer-review

62 Citations (Scopus)

Abstract

In the biosynthetic pathway leading to the tropane alkaloid scopolamine, which is the 6,7β-epoxide of hyoscyamine, the enzyme hyoscyamine 6-hydroxylase (H6H; EC 1.14.11.11) catalyses two consecutive steps, hydroxylation of hyoscyamine followed by epoxide formation, to produce scopolamine. The h6h gene encoding H6H from Hyoscyamus niger, placed under the regulation of the CaMV 35S promoter, was introduced into the genome of a scopolamine-rich Duboisia hybrid by a binary vector system using the Agrobacterium rhizogenes strain LBA9402. The presence and overexpression of the transgene in the resulting kanamycin-resistant hairy roots and the regenerated plants obtained from them were confirmed by polymerase chain reaction and Northern blot analysis, respectively. The scopolamine levels in the resulting engineered hairy root lines increased up to three times compared to wild-type hairy roots, but there was no clear increase in the engineered regenerated plants. The best engineered line (H6H-5) produced 74.50 mg/l scopolamine, which to our knowledge is the highest content ever reported for hairy roots of tropane-alkaloid-producing plants containing the 35S-h6h transgene.
Original languageEnglish
Pages (from-to)1289-1295
Number of pages7
JournalPlant Science
Volume165
Issue number6
DOIs
Publication statusPublished - 2003
MoE publication typeA1 Journal article-refereed

Fingerprint

Duboisia
scopolamine
Plant Roots
Scopolamine Hydrobromide
Alkaloids
Hyoscyamine
alkaloids
atropine
Tropanes
tropane alkaloids
Genes
Epoxy Compounds
hyoscyamine (6S)-dioxygenase
Transgenes
epoxides
genes
transgenes
Hyoscyamus
Hyoscyamus niger
Agrobacterium

Keywords

  • hairy roots
  • duboisia hybrid
  • tropane alkaloids
  • scopolamine
  • metabolic engineering
  • hyoscyamine
  • hyoscyamine-6beta-hydroxylase

Cite this

Palazon, Javier ; Moyano, Elisabet ; Cusido, Rosa M. ; Bonfill, M. ; Oksman-Caldentey, Kirsi-Marja ; Pinol, M. Teresa. / Alkaloid production in Duboisia hybrid hairy roots and plants overexpressing the h6h gene. In: Plant Science. 2003 ; Vol. 165, No. 6. pp. 1289-1295.
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abstract = "In the biosynthetic pathway leading to the tropane alkaloid scopolamine, which is the 6,7β-epoxide of hyoscyamine, the enzyme hyoscyamine 6-hydroxylase (H6H; EC 1.14.11.11) catalyses two consecutive steps, hydroxylation of hyoscyamine followed by epoxide formation, to produce scopolamine. The h6h gene encoding H6H from Hyoscyamus niger, placed under the regulation of the CaMV 35S promoter, was introduced into the genome of a scopolamine-rich Duboisia hybrid by a binary vector system using the Agrobacterium rhizogenes strain LBA9402. The presence and overexpression of the transgene in the resulting kanamycin-resistant hairy roots and the regenerated plants obtained from them were confirmed by polymerase chain reaction and Northern blot analysis, respectively. The scopolamine levels in the resulting engineered hairy root lines increased up to three times compared to wild-type hairy roots, but there was no clear increase in the engineered regenerated plants. The best engineered line (H6H-5) produced 74.50 mg/l scopolamine, which to our knowledge is the highest content ever reported for hairy roots of tropane-alkaloid-producing plants containing the 35S-h6h transgene.",
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author = "Javier Palazon and Elisabet Moyano and Cusido, {Rosa M.} and M. Bonfill and Kirsi-Marja Oksman-Caldentey and Pinol, {M. Teresa}",
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Alkaloid production in Duboisia hybrid hairy roots and plants overexpressing the h6h gene. / Palazon, Javier; Moyano, Elisabet; Cusido, Rosa M.; Bonfill, M.; Oksman-Caldentey, Kirsi-Marja; Pinol, M. Teresa (Corresponding Author).

In: Plant Science, Vol. 165, No. 6, 2003, p. 1289-1295.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Alkaloid production in Duboisia hybrid hairy roots and plants overexpressing the h6h gene

AU - Palazon, Javier

AU - Moyano, Elisabet

AU - Cusido, Rosa M.

AU - Bonfill, M.

AU - Oksman-Caldentey, Kirsi-Marja

AU - Pinol, M. Teresa

PY - 2003

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N2 - In the biosynthetic pathway leading to the tropane alkaloid scopolamine, which is the 6,7β-epoxide of hyoscyamine, the enzyme hyoscyamine 6-hydroxylase (H6H; EC 1.14.11.11) catalyses two consecutive steps, hydroxylation of hyoscyamine followed by epoxide formation, to produce scopolamine. The h6h gene encoding H6H from Hyoscyamus niger, placed under the regulation of the CaMV 35S promoter, was introduced into the genome of a scopolamine-rich Duboisia hybrid by a binary vector system using the Agrobacterium rhizogenes strain LBA9402. The presence and overexpression of the transgene in the resulting kanamycin-resistant hairy roots and the regenerated plants obtained from them were confirmed by polymerase chain reaction and Northern blot analysis, respectively. The scopolamine levels in the resulting engineered hairy root lines increased up to three times compared to wild-type hairy roots, but there was no clear increase in the engineered regenerated plants. The best engineered line (H6H-5) produced 74.50 mg/l scopolamine, which to our knowledge is the highest content ever reported for hairy roots of tropane-alkaloid-producing plants containing the 35S-h6h transgene.

AB - In the biosynthetic pathway leading to the tropane alkaloid scopolamine, which is the 6,7β-epoxide of hyoscyamine, the enzyme hyoscyamine 6-hydroxylase (H6H; EC 1.14.11.11) catalyses two consecutive steps, hydroxylation of hyoscyamine followed by epoxide formation, to produce scopolamine. The h6h gene encoding H6H from Hyoscyamus niger, placed under the regulation of the CaMV 35S promoter, was introduced into the genome of a scopolamine-rich Duboisia hybrid by a binary vector system using the Agrobacterium rhizogenes strain LBA9402. The presence and overexpression of the transgene in the resulting kanamycin-resistant hairy roots and the regenerated plants obtained from them were confirmed by polymerase chain reaction and Northern blot analysis, respectively. The scopolamine levels in the resulting engineered hairy root lines increased up to three times compared to wild-type hairy roots, but there was no clear increase in the engineered regenerated plants. The best engineered line (H6H-5) produced 74.50 mg/l scopolamine, which to our knowledge is the highest content ever reported for hairy roots of tropane-alkaloid-producing plants containing the 35S-h6h transgene.

KW - hairy roots

KW - duboisia hybrid

KW - tropane alkaloids

KW - scopolamine

KW - metabolic engineering

KW - hyoscyamine

KW - hyoscyamine-6beta-hydroxylase

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DO - 10.1016/S0168-9452(03)00340-6

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SP - 1289

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JO - Plant Science

JF - Plant Science

SN - 0168-9452

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