• KSBB Journal
• /
• 제15권6호
• /
• pp.609-614
• /
• 2000

최근 항암제로서 관심을 끌고있는 camptothecin올 생산하기 위한 희수(Camptotheca acuminata) 현탁세포 배양과 cam­p ptothecin의 생산을 촉진하는 연구를 수행하였다. 장기간 배양 으로 발생되는 이차대사산물 생산능력의 변이가 관찰되어 이 를 극복하고자 세포의 응집을 통한 camptothecin의 생산을 촉진하고자 하였다. 세포의 응집은 hybrid 배지와 sucrose 4 %일 때에 가장 많이 유도되었고, 이때 camptothecin의 생산량도 $18.04{\times}10^{-4} mg/L$로 가장 높은 생산성을 가졌다. 또한, 세포의 응집을 유도하기 위한 일환으로 교반속도를 변화시켜 이에 대한 효과를 알아보았는데, 100rpm에서 가장 많은 세포응집이 유도되었으며, camptothecin의 생산 역시 $19.4{\times}10^{-4} mg/L$로 가장 높은 결과를 나타내었다.

• Journal of Microbiology and Biotechnology
• /
• 제10권3호
• /
• pp.327-332
• /
• 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
• /
• 제47권1호
• /
• pp.135-140
• /
• 2004

현재 시판되고 있는 한약재 및 약용식물 118종에 대해서 80% 메탄올 추출물을 제조하였고 , 이들의 항산화 활성을 DPPH 와 superoxide anion radical 소거능 측정 방법을 이용하여 조사하였다. DPPH radical 소거 활성은 괴화 (Sophora japonica: 줄기부)의 메탄올 추출물에서 76.9%의 활성을 보였고 회수(Camptotheca acuminata Dence: 지상부)의 추출물에서는 50.9%의 활성을 보였으며 박하(Mentha arvensis: 지상부), 비파엽 (Eriobotryajaponica: 지상부), 소엽(Perilla frutescens: 줄기부), 애엽(Artemisia asiatica: 줄기부 ), 초과(Amomum. costatum: 지상부) 의 추출물과 산비장이(Serratula koreana: 잎), 살구나무(Prunus ansu: 잎)는 30% 이상의 활성을 보였다. Superoxide anion radical 소거능은 박하(Mentha arvensis), 비파엽(Eriobotrya jαponica), 초과(Amomum costatum), 희수(Camptotheca acuminata Dence)의 지상부에서 80% 이상의 활성을 보였으며 괴화(Sophora jαponica), 소엽(Perilla frutescens), 애엽(Artemisia asiatica), 순비기나무 (Vitex rotundifolia)의 줄기부 , 짚신나물(Agrimonia pilosa)의 잎, 창질경이(Plantago lanceolata) 지상부는 50% 이상의 활성을 나타내었다. 박하(Mentha arvensis) 와 비파엽(Eriobotrya jαponica), 초과(Amomum costatum), 희수(Camptotheca acuminata Dence)의 지상 부는 DPPH뿐만 아니라 superoxide anion radical 소거능에서도 높은 활성을 보였다.

• KSBB Journal
• /
• 제15권5호
• /
• pp.428-433
• /
• 2000

희수 (Camptotheca acuminata) 현닥세포배양에서 최근 항암 제로서 관심을 팔고있는 camptothecin을 얻기 위하여 연구를 수행하였다. 현탁세포배양에서 exponential phase의 최고 생장 속도는 $0.269day^{-1}$이었으며 배가시간이 2.58일이었다. Camptothecin 은 배양초기에 생성되지 않고 exponential phase 말기부터 서서히 생성되어 stationary phase 초기에 최고를 나타내고 이후 급격히 감소하였다. Camptothecin의 생산을 촉진하기 위하여 elicitor를 사F용하였다. 여러 elicitor 중 jasmonic acid와 cellulase 가 camptothecin의 생성을 증가시켰다, Elicitor는 접종과 동시 에 투여 했을 때 가장 효과가 좋았으며, jasmonic acid와 cellulase 의 combination으로 효과가 더욱 증진했는데 대조구에 비하여 약 20배 정도 증가한 $20.42{\times}10^{-3}mg/\ell$ 의 camptothecin이 생 성되었다. 이것은 생산량을 증가시킨 것뿐만 아니라 생산기 간도 약 1/4로 단축시키는 결과를 얻을 수 있었다.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제3권2호
• /
• pp.91-95
• /
• 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
• /
• 제8권1호
• /
• pp.32-36
• /
• 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.

• 한국약용작물학회지
• /
• 제17권3호
• /
• pp.192-197
• /
• 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
• /
• 제8권6호
• /
• pp.631-638
• /
• 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
• /
• 제16권1호
• /
• pp.66-70
• /
• 2001

순수한 초임계 이산화탄소를 이용하여 camptothecin(CPT)와 10-hydroxy camptothecin(HCPT) 추출 시험을 결과, 이들은 각각 유기용매 메탄올 추출에 비교하여 3%와 2% 밖에 추출되지 않았다. 보조용매로 메탄올과 에탄올을 이용하여 CPT와 HCPT를 추출하면 추출 효율이 상승됨을 확인할 수 있었다. 특히 에탄올 보다 메탄올이 효율이 좋았다. 따라서 보조용매 적용에 관련된 여러 조건들을 최적화 하였다. 메탄올을 보조용매로 사용하는 경우에 보조용매의 농도가 증가할수록 추출 효율도 증가하였고 16% 이상에서는 추출 효율이 완만하게 증가하였다. 또한 추출은 온도에 따라 크게 영향을 받지 않아으나 온도에 따라 약간 증가하는 경향을 보였다. 초임계 추출에서 압력의 영향을 살펴본 결과 250 bar까지 압력이 증가할수록 추출 효율은 증가하였다. Acetate ion을 공급하여 추출 실험을 수행하였으나 CPT나 HCPT 모두 ion pairing agent의 영향은 없었다. 본 시스템과 같은 solid/fluid 추출 시스템에서 추출 효율에 영향을 크게 미치는 3가지 요소 (용해도, 확산, 및 matrix)중 확산의 영향을 살펴보았다. 여러 기하학적 모델 중 matrix가 구형인 hot-ball model로 확산게수를 계산하였다. 본 시스템에서 보조용매를 포함한 초임계 유체의 $D/r^2$ 값은 0.0072 $min^{-1}$로서 순수한 메탄올 유기용매 추출보다 큰 값을 보였다.

• 농업생명과학연구
• /
• 제45권1호
• /
• pp.33-40
• /
• 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.