• Korean Journal of Medicinal Crop Science
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• v.12 no.5
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• pp.366-371
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• 2004

Aconitine alkaloids produced from cell suspension cultures of Aconitum napellus for the first time. The effects of various culture conditions on cell biomass and aconitine accumulation in cell suspension cultures were investigated. Suspension cell cultures of A. napellus were established by transferring callus tissues from leaf explants onto liquid MS medium supplemented with $1\;mg/l$ NAA and $0.1\;mg/l$ kinetin. Among the culture media tested, MS medium had a pronounced effect on cell growth and aconitine accumulation. The maximum dry cell weight was obtained at inoculum size of 3 g (FCW) per flask and in MS medium supplemented with 5% sucrose after 8 weeks. The addition of salicylic acid (SA) and yeast extract (YE) in the MS medium enhanced aconitine accumulation. Using a proper combination of culture condition and supplements, aconitine content could reach 0.043% (dry weight basis), that was $2.5{\sim}3$ fold higher that detected in control cultures.

• KSBB Journal
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• v.13 no.1
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• pp.26-30
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• 1998

The suspension cultures of Mentha piperita produce menthol which has very low solubility in water due to its hydrophobicity. This can be considered as a factor for its low production in the suspension suspension cultures. Cyclodextrin has the hydrophobic cavity inside the molecule in which menthol can be captured and allow to form a stable complex. The suspension culture of Mentha piperita showed 70% higher production enhancement in the medium containing 1.5%(w/v) $\beta$-cyclodextrin than the control. $\beta$-cyclodextrin had no adverse effect on the cell growth and showed the best result among $\alpha$-, $\beta$- and $\gamma$-cyclodextrins tested in terms of menthol production. We demonstrated that $\beta$-cyclodextrin can be used to enhance the production of menthol in the suspension cultures by capturing hydrophobic menthol into the cavity of cyclodextrin molecules.

• Natural Product Sciences
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• v.3 no.1
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• pp.71-74
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• 1997

Cell suspension cultures of Solanum mammosum transformed inoculated salicyl alcohol into salicin $(salicyl\;alcohol\;2-0-{\beta}-D-glucopyranoside)$. The highest level of salicin (59.3 mg/flask) in the cells was formed within 3 days after inoculating with salicyl alcohol (50 mg /flask containing 50 ml medium). The glucosylation capability of salicyl alcohol by cell suspension cultures of S. mammosum was relatively higher than that reported previously.

• Korean Journal of Medicinal Crop Science
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• v.12 no.6
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• pp.442-447
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• 2004

Cell growth and gomisin J production by suspension cultures of Schisandra chinensis Baillon were investigated under various culture media, initial sucrose concentrations, shaking speeds, and inoculum sizes. Callus was induced from in vitro cultivated leaf segments on MS medium supplemented with $1\;mg/{\ell}$ NAA. The maximum dry cell weight of 2.23 g was obtained at inoculum size of 0.5 g fresh cell weight and in MB5 medium supplemented with $1\;mg/{\ell}$ NAA, 3% sucrose after 8 weeks. The production of gomisin J in suspension cell cultures was maximized in WPM medium containing 5% sucrose. The shaking speed for maintaining maximal cell dry weight was 100 rpm while the best shaking speed for gomisin J accumulation was 140 rpm.

• Plant Biotechnology Reports
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• v.5 no.2
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• pp.127-134
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• 2011

The biotransformation potential of cell suspension cultures generated from Withania somnifera leaf was investigated, using withanolides, i.e. withanolide A, withaferin A, and withanone as precursor substrates. Interestingly, the cell suspension cultures showed inter-conversion of withanolides, as well converted to some unknown compounds, released to the culture media. The bio-catalyzed withanolide was detected and quantified by TLC and HPLC, respectively. There is noticeable conversion of withanolide A to withanone, and vice versa though at a lower level. The type of reaction of this biotransformation appears to be substitution of 20-OH group to 17-OH in withanolide A. In this paper, we present for the first time the possibility of biotransformation by inter-conversion of withanolides of pharmacological importance through cell suspension culture of W. somnifera. The possible role of putative cytochrome $P_{450}$ hydroxylases is implicated in the conversion.

• Journal of Plant Biotechnology
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• v.6 no.2
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• pp.125-130
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• 2004

Callus and cell suspension cultures of Eurycoma longifolia Jack could be an alternative supply of 9-hydroxycanthin-6-one and 9-methoxycanthin-6-one. The callus tissues were initiated from leaves of different trees. The friable calli were used for the preparation of the cell suspension cultures of E. longifolia. The leaf explant of tree Eu-9 produced the most callus and also induced high cell biomass in the cell suspension culture, but it produced low quantity of 9-methoxycanthin- 6-one and 9-hydroxycanthin-6-one. The leaf explant from tree Eu-8 produced low quantity of callus and cell biomass, but produced the highest quantity of 9-methoxycanthin- 6-one and 9-hydroxycanthin-6-one. Optimum production of cell biomass was obtained on cell culture medium with pH 5.75 prior to autoclaving, but high alkaloids content could be induced in culture medium in acidic condition with pH 4.75 and 5.25 prior to autoclaving.

• Journal of Microbiology and Biotechnology
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• v.8 no.6
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• pp.631-638
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• 1998

Studies were made to elucidate the cell growth and the production of camptothecin and its derivatives in cell cultures of Camptotheca acuminata. High resolution HPLC chromatograms to analyze camptothecin and 10-hydroxycamptothecin in lactone and carboxylate forms were obtained with a fluorescence detector. Calli inductions were optimized with the young stem of explant on Schenk and Hildebrandt (SH) medium supplemented with 5 mg/l $\alpha$-naphthaleneacetic acid (NAA), 0.2 mg/l 6-benzylamino purine (BAP), 2.0% sucrose, and 0.5% agar. The hybrid medium, a mixture of SH and Murashige and Skoog (MS) salts, was developed for homogeneous suspension cultures without large cell aggregates. The optimum phytohormone concentrations for successful suspension cultures were 1.0mg/l of 2,4-D and 0.5 mg/l of kinetin. The highest growth in suspension cultures was observed when 49.7% (w/w) of the cells was composed of small aggregates which were below 0.1 mm in diameter. Time course changes of cell growth and camptothecin production showed that camptothecin accumulation was started at the end of the growth phase and the maximum content was obtained 10 days after inoculation. Yeast extract elicitor increased camptothecin accumulation 4 times. Methyl jasmonate and jasmonic acid also increased camptothecin production 6 and 11 times, respectively.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.198-204
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• 2005

Seeds of neem were collected from different parts of India and analyzed for their azadirachtin content by High Performance Liquid Chromatography (HPLC). In order to assess the effects of genotypic and geographical variation on azadirachtin content in cell cultures, callus development was attempted in the seeds containing high and low concentration of azadirachtin. The concentration of azadirachtin in callus cultures was significantly affected by the explant source. Seed kernels with higher azadirachtin content produced higher azadirachtin content in callus cultures and lower azadirachtin content was seen in callus cultures produced from seed kernels with low azadirachtin content. The protocol for development of elite stock culture of Azadirachta indica was established with the objective of selecting a high azadirachtin-producing cell line. The highest azadirachtin-producing cell line was selected and the effects of different media and illumination conditions on growth and azadirachtin production were studied in shake flask suspension culture. Detailed batch growth kinetics was also established. These studies provided elite starter culture and associated protocols for cultivation of A. indica plant cell culture in the bioreactor.

• Korean Journal of Plant Tissue Culture
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• v.26 no.4
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• pp.289-291
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• 1999

Hypocotyl explants from 7 days old seedlings of one $F_1$ hybrid cultivar and two pure lines of cucumber formed embryogenic calli at frequencies of up to 8% when cultured on Murashige and Skoog medium (MS) supplemented with 1 mg/L 2,4-D for 3 weeks. Embryogenic calli gave rise to somatic embryos. When slices of somatic embryos were cultured on the same medium for 4 weeks, they formed embryogenic calli. Embryogenic cell suspension cultures were established with embryogenic calli in MS liquid medium with 1 mg/L 2,4-D. Embryogenic potential of cell suspension cultures was maintained by subculturing every seven days. When the level of 2,4-D in the medium was lowered to 0.2 mg/L by diluting with liquid MS basal medium, embryogenic cell suspension cultures underwent development into numerous somatic embryos. When plated onto MS basal medium, over 95% of somatic embryos developed into plantlets. Plantlets were transplanted to potting soil and grown to maturity.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.6
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• pp.331-334
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• 2002

Culture conductivity and on-line NADH fluorescence were used to measure cellular growth in plant cell suspension cultures of Podophyllum hexandrum. An inverse correlation between dry cell weight and medium conductivity was observed during shake flask cultivation. A linear relationship between dry cell weight and culture NADH fluorescence was obtained during the exponential phase of batch cultivation In a bioreactor under the pH stat (pH 6) conditions. It was observed that conductivity measurement were suitable for biomass characterisation under highly dynamic uncontrolled shake flask cultivation conditions. However, if the acid/alkali feeding is done for pH control the conductivity measurement could not be applied. On the other hand the NADH fluorescence measurement allowed online-in situ biomass monitoring of rather heterogenous plant cell suspension cultures in bioreactor even under the most desirable pH stat conditions.