• KSBB Journal
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• v.11 no.5
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• pp.600-605
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• 1996

Optimum inoculum concentration for the production of taxol was determined in Taxus brevifolia and Taxus cuspidata cell suspension cultures. By fresh weight, 2.5, 5, 7.5, 10 g/flask of cells were inoculated and cell growth as well as taxol production were examined. In both Taxus cell cultures, the higher the inoculum concentration, the shorter the length of the lag period. The optimum inoculum concentration for taxol production was found to be 5 g/flask. To produce taxol in large quantity, utilization of proper medium was thought to be important. In case of using a production medium with 6% sucrose, taxol production was noticed. Its level reached the maximum at the 9th day of culture and decreased afterwards. However, taxol was not detected from cell cultures in growth medium.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.5
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• pp.350-354
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• 2000

The growth promotion of a Taxus brevifolia cell suspension culture was investigated using conditioning factors. The conditioning factors produced and secreted from cultured cells usually stimulate cell division and the production of secondary metabolites. Therefore, the effective incubation time for the optimal secretion of conditioning factors was firstly determined for the promotion of cell growth. Conditioned media obtained by cultivating for 2 and 5 days showed the promotion of initial cell growth during the early cell growth period. However, the positive effect of the conditioning factors on the initial cell growth did not continue because of the depletion of the medium nutrients. Accordingly, the addition of a carbon source to the conditioned medium prolonged the positive effect on the cell growth. The addition of sucrose to the conditioned medium resulted in the maximum cell density being reached 4 days earlier compared to the control group and an increased substrate yield.

• KSBB Journal
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• v.10 no.2
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• pp.143-148
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• 1995

The effects of various elicitors, metabolic inhibitors and growth regulators on the production of diterpenoid anticancer agent taxol were investigated in cell suspension cultures of Taxus brevifolia. Cell cultures of T. brevifolia were treated by 5 kinds of biotic elicitors, 5 kinds of abiotic elicitors, 2 kinds of metabolic inhibitors and 8 kinds of growth regulators at the end of exponential growth phase. Among those treatments, chlorocholine chloride-an inhibitor of plant steroid metabolism-increased the taxol production most significantly. From a series of optimization studies, it was found that the addition of 1mM of chlorocholine chloride at the 9th day of culture was the best for taxol production. Taxol yield under this condition was 0.72mg/$\ell$.

• Korean Journal of Pharmacognosy
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• v.21 no.2
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• pp.142-147
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• 1990

Yew cortex, twigs and leaves(Florida yew, Taxus floridana Nutt et Champ; Pacific, yew Taxus brevifolia Nutt(Taxaceae)) were extracted with MeOH and $CHCI_3$, respectively, and then the chloroform extract was fractionated by silica column chromatography. As biologically active substances, taxol derivatives were identified and several biological activity tests were followed. The results showed that it bad no antimosquito activity and inhibited the growth of root meristem of cress seeds. The chloroform ext. of Florida yew showed 91%, inhibition rate at 400 ppm and the taxol fractions of Pacific yew did 91% at 200 ppm, 94% at 100 ppm. In the antitumor activity tests in vitro against a cultured cell line of L1210 mouse leukemia, Pacific yew showed 99% inhibition rate at 1.0 ppm and Florida yew did 97% at 10 ppm.

• BMB Reports
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• v.38 no.6
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• pp.668-675
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• 2005

A full-length cDNA encoding taxadiene synthase (designated as TmTXS), which catalyzes the first committed step in the Taxol biosynthetic pathway, was isolated from young leaves of Taxus media by rapid amplification of cDNA ends (RACE). The full-length cDNA of TmTXS had a 2586 bp open reading frame (ORF) encoding a protein of 862 amino acid residues. The deduced protein had isoelectric point (pI) of 5.32 and a calculated molecular weight of about 98 kDa, similar to previously cloned diterpene cyclases from other Taxus species such as T. brevifolia and T. chinenisis. Sequence comparison analysis showed that TmTXS had high similarity with other members of terpene synthase family of plant origin. Tissue expression pattern analysis revealed that TmTXS expressed strongly in leaves, weak in stems and no expression could be detected in fruits. This is the first report on the mRNA expression profile of genes encoding key enzymes involved in Taxol biosynthetic pathway in different tissues of Taxus plants. Phylogenetic tree analysis showed that TmTXS had closest relationship with taxadiene synthase from T. baccata followed by those from T. chinenisis and T. brevifolia. Expression profiles revealed by RT-PCR under different chemical elicitor treatments such as methyl jasmonate (MJ), silver nitrate (SN) and ammonium ceric sulphate (ACS) were also compared for the first time, and the results revealed that expression of TmTXS was all induced by the tested three treatments and the induction effect by MJ was the strongest, implying that TmTXS was high elicitor responsive.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.413.3-414
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• 2002

Paclitaxel is a diterpenoid isolated from Taxus brevifolia and is an active anticancer drug for the treatment of ovarian cancer, breast cancer and Kaposi's sarcoma. Due to its low solubility in water, it is dissolved in Cremophor EL(polyethoxylated castor oil) and ethanol, which cause serious side effects including hypersensitivity. BLK460 was developed as a novel polymeric micellar paclitaxel formulation containing Aceporol460 as solubilizer (omitted)

• KSBB Journal
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• v.11 no.3
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• pp.323-328
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• 1996

Effects of three kinds of precursors as well as exogenous taxanes on the production of taxol and its derivatives were investigated in Taxus cuspidata and Taxus brevifolia cell suspension cultures. When geraniol was added as a precursor of geranylgeranyl pyrophosphate to enhance diterpenoid metabolism, production of some taxanes including taxol was enhanced. The time of addition and amount of feeding were found to be important factors. Feeding of camphor and menthol resulted in negative effects on taxol production. Influences of exogenous taxanes on taxane production by cell cultures were found to be very complicated. When taxol, baccatin III, cephalomannine were exogenously added into the culture, production of baccatin III, 7-epi-10-deacetyltaxol, 10-deacetyltaxol was increased respectively.

• Proceedings of the Plant Resources Society of Korea Conference
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• 1999.10a
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• pp.1-45
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• 1999

To date many kinds of compounds have been obtained from plants kingdom as antineoplastic and anti-cancerous agents. However, there is no special type of compounds for ncancer therapy. Various types of substances are effective for various types of cancers and tumors: for instance, alkaloids, lignans, terpenes and steroids etc. Curcumol obtained from Curcuma aromatica was tested and noticed to be effective against cancer of the uterine cervix clinically Oridonin isolated from Rabdosia ssp.is now investigated for clinical trials in China. Moreover, camptothecine isolated from Camptotheca acuminata is also antineoplastic alkaloid, but is very toxic. Chemical modification has been tried to decrease its toxicity. This compound is now using as clinical agent. Harringtonin was investigated as an anticancerous drug in China. Taxol, a compound with a taxane ring isolated from the bark of Taxus brevifolia, has been demonstrated to have substantial anticancer activity in patients with solid tumors refractory standard chemotherapy. Supply of this drug has severely limited full exploration of its antineoplastic potential. Some efforts are continued in National Cancer Institute NCI) Washington for surveying various Taxus species for optimal taxol content, improvement in semi-synthesis from baccatin III, improvement in method of extraction, and development of alternative renewable resources. Further, there are many compounds which have been reported as antineoplastic agents. On the other hand, we have screened on higher plants collected in Japan, China, Korea, Southeast Asia and South America for antineoplastic activity, which has been done using Sarcoma 180 ascites in mice, P388 Iymphocytic leukemia in mice, Chinese hamster lung V-79 cells, P388 cells and nasopharynx carcinoma (KB) cells in our laboratory, as primary screening. In this meeting, I will present on antitumor and cytotoxic substances of the higher plants (Rubia cordifolia, Ailanfhus Vilmoriniana, Aster tataricus, Taxus cuspidata var. nana, Cephalotaxus harringtonia var drupacea, etc.) selected from above screening tests.

• Proceedings of the Korean Society of Toxicology Conference
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• 2002.05a
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• pp.117-117
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• 2002

Paclitaxel isolated from the pacific yew tree, Taxus brevifolia, is microtuble-stabilizing agent that has a promising anticancer activity against a wide variety of tumors such as ovarian, breast and lung cancers. Because of its poor water solubility, paclitaxel is currently formulated in a mixture of polyoxyethyleneglycerol triricinoleate 35 (Cremophor EL) and dehydrated ethanol USP (1:1 v/v).(omitted)

• BMB Reports
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• v.45 no.10
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• pp.589-594
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• 2012

Paclitaxel is produced by various species of yew trees and has been extensively used to treat tumors. In our research, a taxadiene synthase (TS) gene from Taxus brevifolia was used to transform the roots of cultured ginseng (Panax ginseng C.A. Meyer) to produce taxadiene, the unique skeletal precursor to taxol. The TS gene was successfully introduced into the ginseng genome, and the de novo formation of taxadiene was identified by mass spectroscopy profiling. Without any change in phenotypes or growth difference in a TS-transgenic ginseng line, the transgenic TSS3-2 line accumulated $9.1{\mu}g$ taxadiene per gram of dry weight. In response to the treatment of methyl jasmonate for 3 or 6 days, the accumulation was 14.6 and $15.9{\mu}g$ per g of dry weight, respectively. This is the first report of the production of taxadiene by engineering ginseng roots with a taxadiene synthase gene.