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| Induction of Taxus chinensis var. mairei Hairy Roots and Isolation and Purification of Paclitaxel in Hairy Roots |
| WANG Ying-fang, HAN Bin, LI Zhong, JIA Zhen, CHEN Yan-fen, HU Xu-guang, YANG Ze-min |
| Guangdong Pharmaceutical University, Guangzhou 510006, China |
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Abstract Objective: To explore Induction system of Taxus chinensis var. mairei hairy roots and isolation and purification of paclitaxel in hairy roots. Methods: Genetic transformation of Taxus chinensis var. mairei by three different Agrobacterium rhizogenes, the effect of some factors on transformation efficiency were analyzed. Paclitaxel in hairy roots was isolated and purified by supercritical CO2 fluid extraction, silica gel column chromatography and high performance liquid chromatography(HPLC). Results: The induction system of Taxus chinensis var. mairei hairy roots was established. The factors influenced transformation efficiency contain Agrobacterium species of Agrobacterium rhizogenes, types of explants, pro-culture, co-culture time and hormone concentrations in the medium. High voltage paper electrophoresis (HVPE) indicated that crown gall were expressed in Taxus chinensis var. mairei hairy roots. Paclitaxel in hairy roots was successfully separated and purified by supercritical CO2 fluid extraction and silica gel column chromatography. The purity of paclitaxel was 98% by HPLC assay. Conclusion: Taxus chinensis var. mairei hairy roots induced by Agrobacterium rhizogenes can produce paclitaxel, as source of paclitaxel.
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Received: 28 February 2012
Published: 25 July 2012
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