• KSBB Journal
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• v.13 no.2
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• pp.174-180
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• 1998

Enhanced production of (10-deacetyl) baccatin III and related taxanes was observed in suspension cultures of Taxus baccata Pendula. six % of initial glucose and sucrose concentration increased 10-deacetyl baccatin III production 3.5 and 2.5 times, respectively. Methyl jasmonate, as an elicitor, increased taxane production. Time course changes of taxane production after methyl jasmonate addition showed that baccatin II and 10-deacetyl baccatin III were detected first and paclitaxel, 10-deacetyl taxol and cephalomanine were produced in sequence. Feeding experiments with $500{\mu}M$ of benzoic acid increased 10-deacetyl baccatin III production 10 times. Baccatin III production was also increased 8 times by feeding of $500{\mu}M$ of lysine as a precursor.

• KSBB Journal
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• v.11 no.1
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• pp.100-106
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• 1996

The extraction of taxol and baccatin III from the ground needles of Taxus cuspidata were carried out by using supercritical carbon dioxide with and without cosolvents such as ethylacetate and methanol at 300 bar and 313K. Taxol is a promising anticancer agent and baccatin III is a precursor of semisysthesis of taxol. The taxol and baccatin III contents in the extracts were determined by a HPLC. The highest yields of taxol and baccatin III could be obtained by the supercritical extraction with 3wt% methanol and ethylacetate, respectively, as cosolvents. It was also found that the selectivity of taxol and baccatin III were 0.117 and 1.245wt%, respectively, with 0.7wt% ethylacetate, which demonstrated that the selectivity of taxol and baccatin III were increased about 1.8 and 19 times than those of conventional organic solvent extraction.

• YAKHAK HOEJI
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• v.44 no.1
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• pp.60-65
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• 2000

The yeast extract, coconut water, safflower seed oil, arachidonic acid, linolenic acid, jasmonic acid and methyl jasmonate were added to Gamborg's B$_{5}$ medium. The changes on productivity of baccatin III were estimated every 30 minutes and the results were compared using the selected high yielding cell culture system of Taxus cuspidata. In most cases, the peaks of baccatin III productivity occured at 90~120 min after addition of elicitors. Among the compound elicitors, safflower seed oil showed the highest productivity of baccatin III. Also arachidonic acid and linolenic acid increased the baccatin III production.

• KSBB Journal
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• v.11 no.3
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• pp.263-269
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• 1996

Anticancer agent taxol was synthesized from baccatin III which was extracted from cell cultures of European yew, Taxus baccata Pendula. Callus and suspension cultures of T. baccata Pendula showed the formation baccatin III. The content of baccatin III in cell cultures reached 0.015% (w/w) on dry weight basis. The semisynthetic approaches were made with baccatin III and taxol side chain. To prepare taxol side chain, (-)-N-((S)-2-hydroxy-1-phenylethyl) benzamide was synthesized first from (S)-(+)-phenylglycine. Then (-)-N-((IS,2S)-2-hydroxy-1-phenyl-3-butenyl) benzamide was synthesized with vinyl magnesium bromide. The synthesis of (2R, 3S)-(-)-2-(1-ethoxyethoxy)-3-phenyl-3-(phenylmethanamido) propanoic acid with RuCl3, catalyst was the final step to prepare taxol side chain. The semisynthetic reactions yielded 0.0002% taxol, 0.0005% 7-epi-10-deacetyltaxol, and unidentified taxanes on dry weight basis. It is suggested that the semisynthesis of taxol from baccatin III could be an alternative source of taxol and related taxanes.

• Environmental Mutagens and Carcinogens
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• v.16 no.1
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• pp.6-12
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• 1996

To investigate the clastogenicity of taxol and its precursor, 10-aleacetyl baccatin III, we performed chromosomal aberration assay with chinese hamster lung cells in vitro. The IC$_{50}$ values of taxol and 10-deacetyl baccatin III were determined as $1/16 \times 10^{-4}$ M (5.34 $\mu$g/ml) and $1 \times 10^{-2}$ M (560 $\mu$g/ml) in MTT assay, respectively. It means that the cytotoxicity of taxol revealed 100 times more cytotoxic than 10-deacetyl baccatin III in chinese hamster lung cell line. Nevertheless the strong positive genetic toxicity of taxol in the bone marrow micronucleus assay in vivo which was recently reported, we observed weak positive clastogenicity of taxoi only in the absence of metabolic activation system in the concentration ranges used in this experiment. Moreover, to clarify the involvement of metabolic fate of taxol because of its strong positive result in vivo, 10-deacetyl baccatin III which is a precursor in taxol synthesis, also subjected in chromosomal aberration assay in vitro. However, we observed no clastogenicity of 10-deacetyl baccatin III in this experiment. From above results, it was suggested that the esterification at C-13 appears to be relative for its genetic toxicity in chromosome aberration using chinese hamster lung cell in vitro.

• Korean Journal of Pharmacognosy
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• v.29 no.4
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• pp.360-364
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• 1998

To develop new elicitors inducing the high productivity of taxane derivatives, plant growth inhibitors, namely, maleic acid hydrazide, N-phosphomethyl glycine and succinic acid 2.2-dimethyl hydrazide, coconut milk and yeast extract were administrated in the cell suspension culture system of Taxus cuspidata, and the production of baccatin III were analysed. The effects of amino acid related with the biosynthesis of baccatin III were also examined in these culture system. As the results, a remarkable enhancement of baccatin III production was observed in the cultivation with coconut water and with maleic acid hydrazide.

• Korean Journal of Pharmacognosy
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• v.27 no.3
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• pp.262-266
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• 1996

Undifferentiated callus were induced from the young leaves of Taxus cuspidata by treatment with various combinations of plant growth hormones. The effects of light and culturing temperature on production of baccatin III, the precursor of taxol, were studied. The contents of baccatin III in the cultured callus were analysed by HPLC. The illumination of fluorescence and administration of isoleucine, one of the possible substrates in biosynthesis of terpenoids, to the culturing media increased the production of baccetin III.

• KSBB Journal
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• v.14 no.1
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• pp.17-23
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• 1999

Studies were carried out to elucidate the effects of pressure, temperature and mobile phase composition on supercritical $CO_2$ chromatographic separations of paclitaxel, baccatin III, 10-deacetyl baccatin III, 7-epi-10-deacetyltaxol, cephalomanine, and 10-deacetyltaxol. High resolutions of paclitaxel, 10-deacetylbaccatin III, 10-deacetyltaxol were observed with optimized pressure, temperature, and mobile phase composition. The highest resolution between paclitaxel and 10-deacetylbaccain III was observed at 275 kg/$\textrm{cm}^2$, $40^{\circ}C$ with the mobile phase composition of gradient mixture of 3.9-3.6 mL/min $CO_2$, 0.1-0.4 mL/min methanol for 20 min. Resolutions of baccatin III, capalomannine, and 7-epi-10-deacetyltaxol were found to be low in this study. On-line coupled SFE/SFC process was applied to isolate paclitaxel from yew tree powder. As a consequence, paclitaxel with a purity of 95% was obtained with a recovery yield of 38%.

• 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.

• Journal of Microbiology and Biotechnology
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• v.27 no.8
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• pp.1379-1385
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• 2017

The content of taxol in the bark of yews is very low, and this is not affordable from the environmental point of view. Thus, it is a necessity to look for alternative sources of taxol production to solve its supply. Currently, a large portion of the taxol in the market comes from chemical semi-synthesis, but the semi-synthetic precursors such as baccatin III and 10-deacetyl-baccatin III are extracted from needles and twigs of yew trees. Taxol-producing fungi as a renewable resource is a very promising way to increase the scale of taxol production. Our group has obtained a taxol-producing endophytic fungus, Aspergillus niger subsp. taxi HD86-9, to examine if A. niger can produce the taxanes. Six compounds from the fermentation broth of strain HD86-9 were isolated and identified by $^1H$ NMR, $^{13}C$ NMR, and ESI-MS. The results showed that the six compounds included four taxane diterpenoids (taxol, cephalomannine, baccatin III, and 10-deacetyl-baccatin III) and two non-taxane compounds (${\beta}-sitosterol$ and flavonoid isovitexin). The study verified that the taxanes can be produced by the A. niger, which is very important to taxol production via chemical semi-synthesis. Additionally, the finding is potentially very significant to solve the taxol semi-synthetic precursors extracted from needles and twigs of yew trees, and the precursor production can be easily increased through the culture condition optimization, genetic breeding, and metabolic engineering of the A. niger.