• Journal of Microbiology and Biotechnology
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• v.3 no.4
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• pp.238-243
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• 1993

In the suspension cultures of Eschscholtzia californica, phytohormone effects showed that alkaloid production was increased by IAA treatment without kinetin in both volumetric and specific way. Kinetin, however, suppressed alkaloid accumulation. Addition of ethephon inhibited cell growth. However, it enhanced the alkaloid production significantly in both volumetric and specific way. IAA promoted alkaloid production during elicitation. The highest alkaloid accumulation was observed at 5 $\mu$ M of IAA. Ethephon also enhanced alkaloid production during elicitation. The highest alkaloid formation was observed at 460 mg/l of ethephon with elicitation. Elicitation with ethephon, however, altered cell growth and the pattern of benzophenanthridine alkaloids 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.

• 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.

• KSBB Journal
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• v.8 no.5
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• pp.488-494
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• 1993

The accumulation of benzophenanthridine alkaloids, sanguinarine, chelerythrine, chelirubine and macarpine occurred in suspension cultures of Eschscholtzia californica. To increase alkaloid production, feeding experiments with the biosynthetic precursors, tyrosine, tyramine, L-dopa, dopamine with and without elicitation were studied. In feeding experiments with various precursors, the total alkaloid production was slightly increased. The precursor feeding with elicitation, however, increased total alkaloid production several times.

• Korean Journal of Pharmacognosy
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• v.27 no.4
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• pp.289-294
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• 1996

In previous paper, we described the induced callus of Corydalis remota contains a significant amount of alkaloids. This study describes an optimal condition to maximize alkaloid production. The suspension cultures maintained alkaloid production ability after fifth subculture and a small amount of alkaloid seemed to be released out of cells. The yields of alkaloid by cultured cells was varied depending on the concentrations of NAA, carbon sources and phosphate ion and depending on the vitamin combinations and concentrations. Biosynthetic precursor and an elicitor treatment also affected the total alkaloid yield of the cultures. The optimal conditions for alkaloid production were as follows: 1) MS basal salt containing 30 g/l of glucose, 1.0 mg/l of NAA, and vitamins of LS medium should be used. 2) The culture should be treated with tyrosine 20 mg/l, and yeast extract 1.5 ml/l after the culture reached a stationary phase of growth. Five alkaloids were isolated from the cultures and they were characterized. The spectral data unambiguously revealed that the isolated compounds were dihydrosanguinarine, protopine. tetrahydropalmatine, allocyptopine and ambinine, 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.

• 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.

• Journal of Microbiology and Biotechnology
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• v.1 no.3
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• pp.220-226
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• 1991

Large and rapid increases in benzophenanthridine alkaloid production occured in suspension cultures of Eschscholtzia californica cells treated with elicitors. Response to different biotic elicitors showed that elicitors prepared from yeast extract, Collectotrichum lindemuthianum and Verticillium dahliae induced alkaloid formation. Highest alkaloid accumulation was obtained with $60\;\mu\textrm{g}$ of yeast extract elicitor per gram of fresh cell weight. In time course performance after elicitor addition, more than 40 hours were required to obtain saturated alkaloid accumulation. Compounded silicone fluid, an ideal accumulation phase for two-phase culture of E. californica, accumulated a large amount of alkaloids produced in a specific manner. Elicitation in two-phase culture clearly increased net alkaloid production as well as their concentrations in the accumulation phase.

• KSBB Journal
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• v.11 no.4
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• pp.411-419
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• 1996

In this research, an extractive production system for alkaloids, where production and some degree of separation occur simultaneously, was developed in a way that the fast removal of alkaloid produced from the suspension cultures was done by capturing alkaloid with cyclodextrins. The alkaloid production was substantially enhanced up to 40 fold when the solid cultures of E. califonica cells treated with ${\beta}$-cyclodextrin compared to the control. The enhancement of alkaloid production was also observed in the suspension cultures. Interestingly, the production pattern seemed to change when the cultures were treated with ${\beta}$-cyclodextrin so that the major part of the alkaloids in the treated cultures was present in the medium, while the non-treated cultures produced the alkaloids intracellularly. ${\beta}$-cyclodextrin was the most effective one in terms of the alkaloid production among the cyclodextrilns(${\alpha}$-cylodextrin, ${\beta}$-cyclodextrin and ${\gamma}$-cyclodextrin) tested in the suspension cultures. ${\beta}$-cyclodextrin showed no adverse effect on the cell growth. The most effective concentration of ${\beta}$-cyclodextrin was observed around 1.5% (w/v) in the suspension cultures. The formation of the inclusion complex of the alkaloids with ${\beta}$-cyclodextrin in the suspension cultures was confirmed by detecting the shift of UV absorbance from 274 nm to 282 nm with a UV spectrophotometer.

• Journal of Microbiology and Biotechnology
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• v.10 no.1
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• pp.107-110
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• 2000

Elicited cells with methyl jasmonate continued to produce benzophenanthridine alkaloids throughout medium changes in suspension cultures of Eschscholtzia californica. Large increases in alkaloid production were observed by re-elitations with medium changes. The total alkaloid production increased during the successive elicitation steps reaching a maximum level on the 4th elicitation. The highest total alkaloid produced was 250 mg/I, which was 20fold higher than that of the single elicitation and 4-fold higher than that of the normal culture without elicitation. The large increases in alkaloid production in successive re-elicitations with medium changes are believed to be caused by the accumulation of the signal transduction compound, jasmonate.