Taxus media

Paclitaxel was introduced by Bristol-Myers Squibb in 1993. Today, paclitaxel or a precursor can be obtained from cell cultures of Taxus media formed by hybridizing Taxus baccata and Taxus cuspidate. 

T. P. Castor and T. A. Tyler, Determination of taxol in taxus media needles in the presence of interfering components, J. Liquid Chromatogr., 16 123 (1993). 

Efforts to get access to improved manufacturing processes or improved application properties of paclitaxel resulted in two different pathways for the total synthesis of the natural product, as well as analogs. Furthermore, paclitaxel and/or precursors can be obtained under optimized conditions from cell cultures of Taxus media generated by hybridizing Taxus baccata and Taxus cuspidata in overall yields of about 130 mg 11 within two weeks [61]. Paclitaxel can also be obtained by the culture of appropriate microbial strains isolated from paclitaxel producing yew trees [62 - 64]. 

Ketchnm, R.E., Rithner, C.D., Qin, D., Kim, Y.S., WiUiams, R.M., and Croteau, R.B. Taxus metabolomics methyl jasmonate preferentially induces production of taxoids oxygenated at C-13 in Taxus media cell cultures. Phytochemistry 62, 901-909, 2003. Kim, H.-J., Fonseca, J.M., Choi, J.-H., and Kuhota, C. Effect of methyl jasmonate on phenolic compounds and carotenoids of romaine lettuce (Lactuca sativa L.). 7. Agric. Food Chem. 55, 10366-10372, 2007. 

A discovery made by Yukihito Yukimune of Mitsui Petrochemical Industries in 1996 became crucial for achieving higher time-volume yields. [336, 337] He had found that cultivated cells of Taxus media and Taxus baccata produced significantly more paclitaxel and baccatin III, if traces of (-i-/-)-methyl jasmonate were added to the culture medium (Fig. 5.132). 

Cusido RM, Palazon J, Bonfill M, Navia-Osorio A, Morales C, Pinol MT (2002) Improved paclitaxel and baccatin III production in suspension cultures of Taxus media. Biotechnol Prog 18 418 23... 

Exposito O, Syklowska-Baranek K, Moyano E, Onrubia M, Bonfill M, Palazon J, Cusido RM (2010) Metabolic responses of Taxus media transformed cell cultures to the addition of methyl jasmonate. Biotechnol Prog 26 1145-1153... 

Zhang P, Zhou PP, Yu LJ (2009) An endophytic taxol-producing fungus from Taxus media, Cladosporium cladosporioides MD2. Curr Microbiol 59 227-232... 

Wang, C., Wu, J. and Mei, X. (2001) Enhancement of Taxol production and excretion in Taxus chinensis cell culture by fungal elicitation and medium renewal. Applied Microbiology and Biotechnology, 55, 404 410. 

Chitosan oligomer was also used as an elidtor to stimulate the accumulation of secondary metabolites as taxol from suspension cultures of Taxus cuspidate (Li and Tao 2009), anthraquinones in Rubia tinctorum L. (Vasconsuelo et al. 2004). Chitosan from crab shell with DD > 85% was supplemented into cell culture medium of R. tinctorum at a final concentration of 200 mg/L. Chitosan as an elidtor increased in anthraquinone production up to 110% compared to the control. 

In the cell fermentation stage of the process, calluses of a specific Taxus cell line are propagated in a wholly aqueous medium in large fermentation tanks under controlled conditions at ambient temperature and pressure. The feedstock for the cell growth consists of renewable nutrients sugars, amino acids, vitamins, and trace elements. BMS now extracts paclitaxel directly from plant cell cultures, then purifies it by chromatography and isolates it by crystallization. 

Fig. 89.5 Obtaining a cell suspension culture of Taxus baccata for the production of the diterpene alkaloid Taxol. Calli are induced from sterilized segments of the young branches of T. baccata trees. When developed from branch segments, callus pieces are isolated and cultured in solid MS (Murashige and Skoog medium). Friable calli are transferred to liquid medium and cultured in a shaker at 110 rpm in the dark at 25 °C. For details of experimental conditions, see the reference [11]...

The various PLC techniques were compared and trends in PLC summarized by Nyiredy (1996). The following recent applications for compound isolations by classic, capillary-flow PLC have been published photodegradation products of primaquine formed in an aqueous medium (Kristensen et al., 1993) synthesized isoprenoid diphosphates (20 mg of pure compound recovered from a single plate) (Kennedy Keller and Thompson, 1993) brain lipids (Deleva et al., 1992) components from aloe sap (0.75 mm layers of silanized silica gel and 0.5 mm of silica gel were developed in horizontal sandwich chambers) (Wawrzynowicz et al., 1994) taxol and cephalomannine from Taxus cuspidata (silica gel preparative plates developed with heptane-dichloromethane-ethyl acetate, 11 8 1) (Glow-niak et al., 1996) fenvalerate in a pesticide formulation (1-mm silica gel layers developed with hexane-acetone, 9 1, marker spots detected with o-dinitroben-zene-p-nitrobenzaldehyde reagent) (Gupta et al., 1996) and L-omithine-L-... 

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