• KSBB Journal
• /
• v.13 no.2
• /
• pp.174-180
• /
• 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
• /
• v.11 no.3
• /
• pp.263-269
• /
• 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
• /
• v.16 no.1
• /
• pp.6-12
• /
• 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.

• KSBB Journal
• /
• v.14 no.1
• /
• pp.17-23
• /
• 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
• /
• v.11 no.3
• /
• pp.323-328
• /
• 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.

• KSBB Journal
• /
• v.9 no.2
• /
• pp.186-190
• /
• 1994

The content of taxol and related compounds in various tissues of native yews( T. cuspidata var. latifolia) grown in 5 locations of Ullung Island were analyzed. A considerable range of vanation in the content was observed in the needle and bark collected from different trees located at the same area as well as at five different areas. Taxol content was much higher in the needle(0.017% on the dry weight basis) than those in the bark, whereas the content of 10-deacetyl baccatin III(10-DAB III) was slightly higher in the bark(0.073%), regardless of the location of the trees collected. Particuayly, the needle collected from the Hyunpo area, which located in North-west part of Ullung Island, contained the highest level of taxol(0.024%) exceeding the reported level in dried barks of Pacific yew and also somewhat higher level of its Precusor, 10-DAB III(0.049%). These results suggested that the needle of the yew at the Ullung Island could be suitable materials as a renewable source for the mass production of taxol.

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

• Natural Product Sciences
• /
• v.6 no.2
• /
• pp.96-101
• /
• 2000

The ethanolic extract of the heartwood of Taxus baccata L. afforded six taxoids, belonging to the skeletally three different groups, $2{\alpha},\;5{\alpha},\;10{\beta}-triacetoxy-14{\beta}-(2-methyl)-butryloxy-4(20)$,11-taxadiene (1), taxusin (2), baccatin VI (3), baccatin III (4), $1{\beta}-hydroxybaccatin$ I (5) and taxol (6). The isolation and structure elucidation of three taxoids (3-5) have been presented in this study. This is the first report for the presence of these taxoids in Taxus baccata L. growing in Turkey.

• Journal of Applied Biological Chemistry
• /
• v.43 no.3
• /
• pp.176-179
• /
• 2000

Nonpolar taxoides extracted from a large-scale cell culture of Taxus chinensis were isolated through the normal and reverse phase column chromatographies, and their compounds were identified via NMR spectroscopy. The complete separation method was systematically established and described. In dichloromethane, dissolved paclitaxel and other taxoids with hexane were precipitated during the purification of paclitaxel from the plant cell culture of T. chinensis through a large-scale process while the relatively nonpolar taxane derivatives remained dissolved in the hexane phase. 13-Deoxy baccatin III (I), baccatin VI (II), taxchinin I (III), $2{\alpha}$, $5{\alpha}$, $10{\beta}$, $14{\beta}$-tetraacetoxy-4(20), 11-taxadiene(IV), 1-deoxy baccatinVI(V), and taxayuntin C (VI) were isolated through column chromatography and identified via NMR spectroscopy. Compounds I and IV were found to the major components, aside from paclitaxel, in the plant cell culture of T. chinensis. The concentrations of I and IV were compared with the that concentration of the paclitaxel in each of plant cell culture. The possible applications of compounds I, II, IV, and V were discussed.

• Journal of the Korean Wood Science and Technology
• /
• v.27 no.3
• /
• pp.91-98
• /
• 1999

The content of taxol and related compounds in needle and stem tissues of exotic yews growing in the Chollipo Arboretum which located in the West Sea of Korea were determined by HPLC. A considerable variation in contents of taxol and the related compounds within species and among cultivars were observed. The content of taxol was much higher in the needles than those in the young stem tissues except T. X media cv. Vermuleun, T. baccata cv. Washingtonii, and T. X cuspidata cv. Nana, and the content of 10-deacetyl baccatin III(10-DAB III) was also slightly higher in the needle. The needle of T. cuspidata cv. Nigra collected from April was contained the high amount of taxol(1,027 ${\mu}g$ per g dry weight) and also contained high quantity in 10-DAB III(267 ${\mu}g$ per g dry weight). Concentration of taxol in needle was significantly greater than those of meristemic and ovary tissues, Exotic yews growing at the Chollipo Arboretum containing T. cuspidata cv. Nigra could be suitable materials as a source for the tissue and cell cultures for production of taxol.