• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.77-83
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• 1999

We have investigated the effect of culture conditions on tropane alkaloids (scopolamine, hyoscyamine) production in hairy root cultures of Hyoscyamus niger L. induced by Agrobacterium tumefaciens $A_4$T. SH medium was the best for tropane alkaloids production from the hairy root clones, HN18 and HN57. The optimum culture peroid was 5 weeks for HN18 clone and 6 weeks for HN57 clone, respectively. The optimum sucrose and dextrose concentrations in tropane alkaloids productivity were 3% and 2%, respectively. The growth of both HN18 and HN57 clones increased with as sucrose concentration increase up to 7% sucrose, but tropane alkaloid contents was significantly decreased. In the HN18 clone, the optimum concentration of sucrose for alkaloids productivity was 5% and those of dextrose was 2%. The productivity of tropane alkaloids for HN57 clone under dextrose treatments was quite a low level compared to sucrose treatments.

• Korean Journal of Plant Tissue Culture
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• v.25 no.1
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• pp.21-25
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• 1998

The effects of sucrose concentration and some absorbents on growth and tropane alkaloid production in hairy root cultures of Scopolia parviflora were investigated. The maximum effect on growth and tropane alkaloid production in hairy root clone SP11 was obtained for 1/2 B5 medium containing 5% sucrose. The production pattern of tropane alkaloid in hairy roots was some different from that of rhizome of mother plant, particulary showing high littorine contents, which was not found in ordinary roots. Among absorbents examined, charcoal 0.01% and XAD-II 1% made a slight growth promotion effect, whereas the other concentration of charcoal, XAD-II and absorbents (amberlite and chitosan) showed inhibition or no significant effect. The addition of hydroxyapatite enhanced the production of tropane alkaloids significantly than control cultures.

• Korean Journal of Medicinal Crop Science
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• v.11 no.5
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• pp.358-363
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• 2003

ScopoIamine and hyoscyamine which belong to tropane alkaloids are the pharmaceutically valuable anticholinergic drugs. In order to increase the productivities, the effects of elicitation were investigated during hairy root cultures of Scopolia. parviflora. Biotic elicitors originated from 3 fungi and 1 yeast were prepared as homogenate and supernatant and added to 3-week-old cultures. Both of homogenate and supernatant of Candida albicans elicitors increased the scopolamine production. The production of hyoscyamine was enhanced by homogenate of Fusarium solani and supernatant of C. albicans. Most of the other fungal elicitors were also improved on the tropane alkaloid production compared to non-treatment. Among the elicitors tested, C. albicans was proved the optimal biotic elicitor on tropane alkaloids production. These results will be served mass production of tropane alkaloids by large-scale production.

• Korean Journal of Plant Tissue Culture
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• v.24 no.3
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• pp.137-142
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• 1997

In order to elucidate the optimum medium on the growth and tropane alkaloid production of hairy root, hairy root were induced by inoculating leaf and stem of Datura stramonium var. tatula Torr. with Agrobacterium spp. Both Agrobacterium tumefaciens $\textrm{A}_{4}$ T and A, rhizogenes ATCC 15834 among tested strains were effective on hairy root formation. Among 23 clones selected in SH (Schenk and Hildebrandt, 1972) liquid medium, DTLA9 clone was shown fast growth of hairy root and DTLE6 clone was shown high level production of tropane alkaloids. When both DTLA9 and DTLE6 clones were cultured in the GD (Gresshoff and Doy, 1972) medium, alkaloid production was higher than in 8 tested media. It was elucidated that optimum medium for root proliferation and for tropane alkaloid production is SH, GD medium, respectively.

• Korean Journal of Plant Tissue Culture
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• v.24 no.3
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• pp.143-148
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• 1997

The optimum concentrations of pH, sucrose and vitamins for the growth and tropane alkaloid production of hairy root clone DTLA9 (best growth line) were investigated. The optimum pH in growth and tropane alkaliod production of DTLA9 clone in SH (Schenk and Hildebrandt, 1972) basal medium without growth regulator were pH 6.3 and 6.5, respectively. Also, the optimum sucrose concentration in growth and tropane alkaliod production in the same medium were 3.0 and 2.8%, respectively. The optimum concentrations of ascorbic acid, D-pantothenate, nicotinic acid, pyridoxine, riboflavin, and thiamine on the growth of DTLA9 clone in SH basal medium without vitamins were 0.1 mM, 0.003 mM, 0.07 mM, 0.002 mM, 0.025 mM, and 0.01 mM, respectively. In particular, supplement of 0.1 mM ascorbic acid to SH basal medium without vitamins stimulated the tropane alkaloid production.

• KSBB Journal
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• v.17 no.6
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• pp.525-530
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• 2002

The optimum culture conditions for tropane alkaloid production in hairy root cultures of Korea native Scopolia paviflora Nak. were investigated. Hairy root was induced from the rhizome of the mother plant on B5 medium containing 1.0 mg/L IBA. Among the culture media examined, 1/2 B5 medium was the best for tropane alkaloid production, whereas the growth of hairy root increased in SH medium. The best result on the growth of hairy root was obtained in 1.0 mg/L NAA, and tropane alkaloid production was obtained in plant growth regulator-free medium. Of the carbone sources tested, 3% sucrose promoted the growth of hairy root, whereas 5% sucrose increased tropane alkaloid production. Optimum inoculum densities for root growth and tropane alkaloid production were 0.5 g and 1 g, respectively. The addition of XAD resins (1 % w/v) to hairy root cultures led to increases in tropans alkaloid production, and the release of alkaloid into the medium and its adsorption by the resin accounted for about 50 to 80％ of total production. It is concluded that optimized culture conditions and the addition of XAD resins could be used in the development of a bioprocess for tropane alkaloid production in hairy root cultures of S. paviflora Nak.

• Journal of Plant Biotechnology
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• v.29 no.3
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• pp.199-207
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• 2002

The tropane alkaloids hyoscyamine (its racemic form being atropine) and scopolamine are used medicinally as anticholinergic agents that act on the parasympathetic nerve system. Because they differ in their actions on the central nervous system, currently there is a 10-fold higher commercial demand for scopolamine, in the N-butylbromide form, than there is for hyoscyamine and atropine combined. Several solanaceous species have been used as the commercial sources of these alkaloids, but the scopolamine contents in these plants often are much lower than those of hyoscyamine. For this reason there has been long-standing interest in increasing the scopolamine contents of cultivated medicinal plants. Naturally occurring and artificial interspecific hybrids of Duboisia have high scopolamine contents and are cultivated as a commercial source of scopolamine in Australia and other countries. Anther culture combined with conventional interspecific hybridization also has been used to breed high scopolamine-containing plants in the genera Datura and Hyoscyamus, but without much success. The use of recombinant DNA technology for the manipulation of metabolic processes in cells promises to provide important contributions to basic science, agriculture, and medicine. In this review, I introduce on the enzymes and genes involved in tropane alkaloid biosynthesis and current progress in metabolic engineering approaches for tropane alkaloid, especially scopolamine, production.

• YAKHAK HOEJI
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• v.43 no.6
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• pp.729-735
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• 1999

Scopolia rhizome is mistaken as an atractylodes rhizome because of their similarities in shape. That is why atractylodes rhizome imported from China sometimes contain scopolia rhizome, which is very toxic. 8 persons were intoxicated atractylodes after taking imported atractylodes rhizome which is tainted. In kampo medicine prepared with such imported atractylodes rhizome, the level of tropane alkaloids ranged from 1.12∼4.34 mg/dose. In this study, we tried to investigate the tissue distribution of scopolia rhizome in rats. The extracts of scopolia was administered orally to rats (a single dose of 10mg/kg, 20mg/kg and 7 days repeated dose of 10mg/kg). Their blood was collected at 0.5, 1, 2, 4, 6 hrs, and liver, kidney, lung and spleen were collected after 6 hrs. The tissue homogenate was applied to solid phase extraction column for the determination of tropane alkaloids. After the oral administration of 20mg/kg scopolia extracts, l-hyoscyamine was detected in rat blood to 2 hrs after dosing. The concentration of tropane alkaloids was the highest in liver followed by lung, kidney and spleen. However, lung, kidney and spleen were similar in amount.

• Journal of Plant Biology
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• v.36 no.3
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• pp.225-231
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• 1993

Transformed hairy roots of Scopolia parviflora, producing tropane alkaloids and native to Korea, were obtained following infection of rhizome segments with Agrobacterium rhizogenes A4. Each root tip induced from inoculum sites excised and cultured in MS agar or liquid medium. About seventy of hairy root clones were established. Among them, several fast growing hairy root clones were examined for alkaliod content. Two dimensional TLC analysis showed that the tropane alkaloid pattern of hairy root was more complicated than that in the rhizome of mother plant. On the other hand, some hairy root clones did not produce scopolamine and hyoscyamine. In HPLC analysis, some hairy root clones yield higher levels of scopolamine and hyoscyamine than those of mother plant rhizome which used for infection. Scopolamine and hyoscyamine were identified by comparison of their retention times and of their spectra data with those of authentic compounds.

• Korean Journal of Medicinal Crop Science
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• v.12 no.2
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• pp.123-128
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• 2004

Hyoscyamine and scopolamine are two most common tropane alkaloids found in the Solanaceae. The pEB expression vector carrying Nspmt gene was transformed to Agrobacterium rhizogenes. The growth of transgenic hairy roots were approximately to 80% of the wild type root. Transgenic hairy roots are less developed on only their branch roots than those of the wild type root. The extracts from Nspmt transgenic hairy root lines, 3 and 5 contained between 3.52 and 4.23 mg/g dry weight as hyoscyamine and did between 5.23 to 6.40 mg/g dry weight as scopolamine. These results showed that the overexpression of the pmt gene enhanced tropane alkaloids production of S. parviflora transformed roots and this improvement affected both hyoscyamine and scopolamine production.