|Title:||台灣紅豆杉細胞培養生產紅豆杉醇||Other Titles:||Taxol Production by Cell Cultures of Taxus mairei||Authors:||李承榆
|Keywords:||台灣紅豆杉;紅豆杉醇;細胞懸浮培養;生物反應器;癒合組織;Taxus mairei;taxol;cell suspension culture;bio-reactor;callus||Issue Date:||Apr-1995||Publisher:||農業試驗所||Related Publication(s):||農業試驗所特刊第48號||Start page/Pages:||137-148||Source:||臺灣地區藥用植物資源之開發與利用學術研討會專刊||Conference:||臺灣地區藥用植物資源之開發與利用學術研討會
Proceeding of A Symposium on Development and Utilization of Resources of Medicinal Plants in Taiwan
台灣紅豆杉葉片萃取液以HPLC分析結果顯示含有葉片鮮重0.007-0.0267％的紅豆杉醇，以及其他taxanes包括：10-deacetyltaxol、10-deacetylcephalomannine及10-deacetyl-baccatin III。植物細胞及組織培養技術提供生產紅豆杉醇的新方法。本試驗在探討台灣紅豆杉細胞培養生產紅豆杉醇及相關taxanes的技術。台灣紅豆杉的枝條及葉片以添加2mg/l2,4-D或NAA的Gamborg B5培養基培養，約二週後可誘導癒合組織產生。癒合組織繼代培養一個月其重量為原來的五倍。乾物重約為溼重的5-7％。懸浮培養二個月的細胞團可再生出紅豆杉的根。紅豈杉細胞三角瓶懸浮培養液，以RP-18的管柱及配有photo-diede array spec-troscopy的HPLC分析，經三角瓶培養懸浮細胞一個月，可產生紅豆杉醇0.2 mg/l。培養液添加1 mM的phenylalanine，紅豆杉醇含量可達3mg/1。
Needles extract of Taxus mairei was analyzed by high performance liquid chromatography. The taxol content of the extract was about 0.007-0.027 % on the fresh weight basis. Besides taxol. other taxanes ‘such as 10-deacetyltaxol, 10-deacetylcephalomannine and 10-deacetylbaccatin III were also found in the needles extract. Plant cell and tissue culture methods offer a potential alternative for taxol production. This experiment was designed to produce taxol and related taxanes on the cell culture of Taxus mairei. Needles and stems from Taxus mairei were cultured on Gamborg’s B5 medium supplemented with 2ppm 2,4-D or NAA. Callus was induced from the explants after 2 weeks. The biomass of callus increased about 5 times and dry matter content was 5-7% after one month in culture. Root formation was observed from suspension cell after 2 months in culture. Taxol produced by the cell suspension culture was analyzed by HPLC. HPLC analysis was performed on a RP-18 reverse phase column with a photo-diode array spectroscopy. Taxol content in the flask suspension cell was 0.2mg/l after one month cultivation. However, the yield increased to 3mg/l by feeding 1mM phenylalanine to the medium.
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