Solasodine production from self-immobilised Solanum aviculare cells

Tsoulpha, Parthena; Doran, Pauline M.

doi:10.1016/0168-1656(91)90077-9

Cited by 32 publications

(13 citation statements)

References 19 publications

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“…Dry weight was obtained after freeze-drying the cells overnight at -40oC. Steroidal alkaloids in the freeze-dried cells and medium were extracted and analysed against solasodine standards (Sigma, USA) using the methods described previously (Tsoulpha and Doran, 1991). Steroidal-alkaloid levels are quantified as mg solasodine equivalents per g dry biomass.…”

Section: Methodsmentioning

confidence: 99%

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Effect of cell-cycle inhibition on synthesis of steroidal alkaloids by Solanum aviculare plant cells

Mak

Doran

1993

Biotechnol Lett

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Addition of 40 mg 1-1 hydroxyurea to Solanum aviculare cultures resulted in complete inhibition of growth and an increase in the levels of steroidal alkaloids in the biomass. These results demonstrate that solasodine synthesis and growth can be uncoupled, and suggest that steroidal-alkaloid production can be significantly enhanced by inhibiting growth at high cell density. Treatment with 1 rng 1-1 abscisic acid had a negligible effect on growth and product synthesis.

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Section: Methodsmentioning

confidence: 99%

“…In previous work (Tsoulpha and Doran, 1991) we reported the development of smooth, regularly-shaped, 'self-immobilised' aggregates of S. aviculare with diameters up to 2.0 cm. The concentration of steroidal alkaloids in these aggregates was 3 mg g-1 dry weight.…”

Section: Introductionmentioning

confidence: 99%

Effect of cell-cycle inhibition on synthesis of steroidal alkaloids by Solanum aviculare plant cells

Mak

Doran

1993

Biotechnol Lett

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“…Callus was initiated from seedlings and smooth spherical aggregates developed as reported previously (Tsoulpha & Doran 1991). Thirty 250-ml flasks each containing approx.…”

Section: Solanum Aviculare Aggregatesmentioning

confidence: 99%

“…For example, when the linear correlation for dry weight shown in Fig. 8 was used to predict final biomass levels in an air-driven bioreactor (Tsoulpha & Doran 1991), the calculated value was only about 1/3 that actually measured.…”

Section: Atropa Belladonna Hairy Rootsmentioning

confidence: 99%

“…The work is also motivated by recent reports mentioning failure of conductivity to correlate growth of Catharanthus roseus hairy roots (Toivonen et al 1990) and Medicago sativa cell suspensions (McDonald & Jackman 1989). Cell suspensions of Atropa belladonna are examined as a comparatively homogeneous culture; hairy roots of A. belladonna and self-immobilised Solanum aviculare aggregates (Tsoulpha & Doran 1991) provide examples of heterogeneous cultures where conductivity measurement would be especially useful. Relationships between culture conductivity and dry-and fresh-weight changes for these three systems are reported.…”

Section: Introductionmentioning

confidence: 99%

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Limitations associated with conductivity measurement for monitoring growth in plant tissue culture

Kwok

Tsoulpha

Doran

1992

Plant Cell Tiss Organ Cult

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The suitability of conductivity measurement for monitoring growth in plant cell culture has been tested using suspended cells and genetically-transformed hairy roots of Atropa belladonna, and aggregated cells of Solanum aviculare. Other researchers have proposed that a constant ratio exists between increase in cell concentration (Ax) and decrease in medium conductivity (AC). In all cases studied in this work, Ax/AC was not constant over a wide range of cell densities tested in batch culture. With cell suspensions, Ax/AC decreased continuously during the growth phase from 3.4 to 2.5 g cm 1-1 mS-~. For the hairy roots, the ratio between Ax and AC varied by as much as 4-fold during growth. The relationship between conductivity and growth for S. aviculare aggregates was found to vary depending on inoculum density. No simple correlation between conductivity change and cell growth was apparent for the plant-cell systems studied.

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Coculture of genetically transformed roots and shoots for synthesis, translocation, and biotransformation of secondary metabolites

Subroto

Kwok

Hamill

et al. 2000

Biotechnol. Bioeng.

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Genetically transformed shooty teratomas of Atropa belladonna and a Duboisia leichhardtii × D. myoporoides hybrid were studied for biotransformation of tropane alkaloids in shake flasks and bioreactors. Although de novo synthesis of hyoscyamine and scopolamine was limited, shoots of both species were able to translocate and accumulate significant quantities of exogenous alkaloid. The maximum yield of scopolamine from hyoscyamine fed to the Duboisia hybrid shoots was 35% w/w; the yield of the scopolamine precursor, 6β‐hydroxyhyoscyamine, was 37% w/w. Biotransformation activity was poor in A. belladonna shooty teratomas provided with exogenous hyoscyamine; however, scopolamine levels comparable with those in leaves of the whole plant accumulated in shoots fed with hairy root extract. Coculture of A. belladonna shooty teratomas and hairy roots in the same hormone‐free medium was investigated as a means of providing a continuous source of hyoscyamine for conversion to scopolamine. Of the biotransformation systems tested with A. belladonna, coculture produced the highest levels of scopolamine and the highest scopolamine: hyoscyamine ratios. Cocultured shoots accumulated up to 0.84 mg g−1 dry weight scopolamine, or 3–11 times the average concentrations found in leaves of the whole plant. The scopolamine: hyoscyamine ratio in coculture ranged from 0.07 to 1.9, a significant improvement over levels of 0–0.03 normally found in A. belladonna hairy roots. Addition of Pluronic F‐68 or copper sulfate to the medium and variation in initial medium pH did not improve hyoscyamine release from hairy roots. Scopolamine levels were increased using 1 μM copper sulfate or initial medium pH between 6.0 and 8.0; however, results from elicitation of hairy roots could not match the beneficial effect on scopolamine synthesis of root‐shoot coculture. Addition of 0.001–1.0% (w/v) Pluronic F‐68 to the roots reduced hyoscyamine release but postponed necrosis in the root tissue for up to 60 d. © 1996 John Wiley & Sons, Inc.

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Solasodine production from self-immobilised Solanum aviculare cells

Cited by 32 publications

References 19 publications

Effect of cell-cycle inhibition on synthesis of steroidal alkaloids by Solanum aviculare plant cells

Effect of cell-cycle inhibition on synthesis of steroidal alkaloids by Solanum aviculare plant cells

Limitations associated with conductivity measurement for monitoring growth in plant tissue culture

Coculture of genetically transformed roots and shoots for synthesis, translocation, and biotransformation of secondary metabolites

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