• KSBB Journal
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• v.15 no.6
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• pp.609-614
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• 2000

Suspension cultures of Camptotheca acuminata, which is known to produce the anticancer indole alkaloid camptothecin and its derivatives, were made to increase camptothecin production. The capability of camptothecin production in suspended cells is decreased by repeated subculturein. Aggregated cells produced more camptothecin than single cells. Optimal cell aggregation was achieved in hybrid medium supplemented with 4% sucrose. Aggregated cells in hybrid medium with 4% sucrose produced $18.04{\times}10^{-4} mg/L$ of camptothecin. The control of shaking speeds was effective at inducing cell aggregation and camptothecin production. A shaking speed of 100 rpm was found optimum to increase the cell aggregation with a camptothecin production of $19.4{\times}10^{-4} mg/L$.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.327-332
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• 2000

Supended cells of Camptotheca acuminata were observed to lose their ability to synthesize camptothecin and its derivatives as a result of repeated cultures. Accordingly, the maintenance of high-yield cells by cryopreservation was sudied to overcome this stability problem, and various factors involved were optimized. Pregrowing the cells in 8% myoinositol for 4 days was found to be the most effective in improving survival. The highest survival was obtained when the pregrown cells were cryoprotected with a mixture of 10% DMSO, 0.6M mannitol, and 10% glycerol. When the cryopreserved cells were maintained in a freezer at $-70^{\circ}C$, 94% survival was obtained after 4 months. The survivals after 5 and 8 months of storage decreased to 52% and 45%, respectively. No loss of biosynthetic capacility of camptothecin was observed after short to medium term cryopreservation of C. acuminata.

• Applied Biological Chemistry
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• v.47 no.1
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• pp.135-140
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• 2004

The antioxidant activities of 80% methanol extracts obtained from 118 medicinal plants were tested through the evaluation of DPPH and superoxide anion radicals scavenging activity. Methanol extracts of Sophora japonica (76.9%) and Camptotheca acuminata Dence (50.9%) were found to have more than 50% DPPH radical scavenging activity while those of Perilla frutescens (37.2%), Amomum costatum (34.9%), Prunus ansu (33.2%), Mentha arvensis (32.3%), Serratula koreana (32.2%), Eriobotrya japonica (30.5%), and Artemisia asiatica (30.5%) showed more than 30% scavenging activity. Even though all of the commercial antioxidants didn't show superoxide anion radical activity, Mentha arvensis (87.7%), Eriobotrya japonica (84.9%), Amomum costatum (82.9%), Camptotheca acuminata Dence (82.1%) showed more than 80% scavenging activity. Mentha arvensis, Eriobotrya japonica, Amomum costatum, Camptotheca acuminata Dence showed strong antioxidative activity in the both DPPH and superoxide anion radical scavenging activities.

• KSBB Journal
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• v.15 no.5
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• pp.428-433
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• 2000

Cell cultures of Camptotheca acuminata, which is known to produce the anticancer indole alkaloid camptothecin and its dervatives, were made to enhance the productivity of camptothecin. In suspension cultures, the maximum cell growth rate in exponential growth phase was $0.269day^{-1}$ which was correlated to 2.58days of cell doubling time. The production of camptothecin was non-growth associated. The camptothecin production was the highest at 11th day form inoculation and then it decreased. Various elicitors were applied to enhance the production of camptothecin. Both of jasmonic acid and cellulase increased the production of camptothecin. The optimal dosing time of elicitor was the beginning of the cultures. The combination of two elicitors was more effective to produce camptothecin than single applications. $20.42{\times}10^{-3}mg/\ell$ of camptothecin was obtained with combined application of two elicitors in two days.

• Biotechnology and Bioprocess Engineering:BBE
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• v.3 no.2
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• pp.91-95
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• 1998

Camptothecin productoin was increased with elicitors, methyl jasmonate, jasmonic acid, yeast extract elicitor, and ferulic acid in suspension cultures of Camptotheca acuminata. Jasmonic acid was found to be the most efficient elicitor. Camptothecin production increased 11 times by using the optimum dosing concentration of jasmonic acid which was 50 ${\mu}$M. The kinetics of camptothecin accumulation in response to the treatment with jasmocin acid showed that the comptothecin accumulation reached the maximum value at 4 days after jasmonic acid dosing and then a rapid decrease in camptothecin accumulation was observed.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.1
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• pp.32-36
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• 2003

Production of camptothecin ((PT) from callus cultures of Camptotheca acuminata Decne was affected by light and culture conditions. Among the culture media tested, modified B5 medium containing 3% (w/v) sucrose, 2 mg/L B,4-D, 2 times of MS medium vitamins, 500 mg/L casein hydrolysate, 250 mg/L myo-inositol, 0.05% (w/v) activated charcoal, and 0.15% (w/v) gelite was used for callus induction . The highest cell growth and CPT production were obtained in dark and green light condition, respectively. Photoperiod has no effect on cell growth and CPT production. Both cell growth and CPT production were also influenced by combination ratio of red and blue light .Cell growth and CPT production were the highest in the ratio of red and blue light,90:10.

• Korean Journal of Medicinal Crop Science
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• v.17 no.3
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• pp.192-197
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• 2009

Camptotheca acuminata, a native of South China is a well known natural source of monoterpene-indole alkaloid camptothecin(CPT), one of the most promising anti-tumoural compounds. This study was conducted to optimize plant growth regulators and culture conditions on plantlets regeneration through organogenesis from callus of Camptotheca acuminta. Callus were induced from various explants of in vitro germinated plantlets of C. acuminta using WPM medium containing 0.2 ㎎/L 2,4-D. Hypocotyl segments were exhibited higher embryogenic callus than the other explants. Shoot buds formation from embryogenic callus was affected by plant growth regulators, pre-treated dark condition and liquid culture. Organogenesis was optimal in WPM liquid medium containing 0.5 ㎎/L BA. The dark pre-treatment for 2 weeks before the solid culture was effective for organogenesis. The regenerated shoots were rooted in WPM medium with 0.2 ㎎/L NAA and successfully acclimated in green-house conditions.

• 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.

• KSBB Journal
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• v.16 no.1
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• pp.66-70
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• 2001

Factors affecting the supercritical carbon dioxide extraction of camptothecin(CPT) and 10-hydroxycamptothecin(HCPT) from the dried powder of Cmptotheca acuminata were studied. Only a few amount of CPT and HCPT was extracted with pure supercritical carbon dioxide. Methanol and ethanol were efficient modifiers to extract CPT and HCPT. At $40^{\circ}C$, 250 bar, 1 mL/min flow rate, 41% of CPT and 35% of HCPT were extracted with supercritical carbon dioxide modified with 16% of methanol. The diffusion effect of HCPT on extraction efficiency was studied in this solid-fluid system. Round matrix hot-ball model assumption revealed that the value of D/$r^2$ was 0.0072 $min^{-1}$ which was higher than that of solvent extraction with methanol.

• Journal of agriculture & life science
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• v.45 no.1
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• pp.33-40
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• 2011

An efficient method for the rapid micropropagation of Camptotheca acuminata from axillary buds was established by application of various plant growth regulators. Among various cytokinins, $0.5mg\;L^{-1}$ BA showed the best performance on shoot multiplication, number average multiple shoots up to 10.8. The propagated shoot cuttings in vitro were elongated on NN basal medium without plant growth regulators. The secondary multiple shoots were induced at the site of initially induced buds. Rooting was induced directly near the base of the shoot on half-strength NN medium containing $0.5mg\;L^{-1}$ of IBA, whereas high concentration of $1.0mgL^{-1}$ IBA could induce callus at the base of the shoot. The camptothecin content, anticancer compound of the micropropagated plants was contained in various tissues. Camptothecin contents were 1.8 and $2.5mg\;g^{-1}$ dry weight in stems from propagated in vitro and mother plant, respectively. This result may be used to develop strategies for large-scale propagation of elite C. acuminata trees.