Combined use of six-well polystyrene plates and thin layer chromatography for rapid development of optimal plant cell culture processes: application to taxane production byTaxus sp. |
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Authors: | V Srinivasan S C Roberts M L Shuler |
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Institution: | (1) School of Chemical Engineering, Cornell University, 14853 Ithaca, NY, USA |
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Abstract: | TwoTaxus (T. chinensis andT. baccata) cell suspension cultures were used as a model system to demonstrate the similarities of biomass accumulation and secondary metabolite (taxane) production obtained from cultures in six-well polystyrene plates and glass shake flasks (25 ml and 125 ml). Interference from binding of taxanes in cell-free culture broth to the polystyrene plates was minimal with 85% of the paclitaxel (Taxol®) and 100% of baccatin and 10-deacetyl-7-xylosyl-taxol remaining in the medium after 24 h beyond which no further binding was observed. A simple thin layer chromatography (TLC) procedure with a chloroform: acentonitrile (4:1) solvent system on silica gel was developed to simultaneously test up to 17 cultures for taxane production. The combination of six-well plate technology for experimentation and TLC for rapid taxane analysis can greatly accelerate the establishment of conditions for an optimalTaxus plant-cell culture process for taxane production.Abbreviations
TLC
Thin layer chromatography
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2,4D
2,4-dichlorophenoxyacetic acid
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HPLC
high pressure liquid chromotography
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UV
ultraviolet
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Rf
retention factor |
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Keywords: | Taxus Taxol Paclitaxel Taxane TLC Secondary metabolite Plant cell cultures Suspensions Six-well plate |
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