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为利用现代生物技术生产紫杉醇,研究细胞悬浮培养过程中紫杉醇的生物合成,研究了细胞生长与培养液中主要营养物质的消耗情况。在Gamborg’sB5培养液中加入诱导子,对本实验室诱导筛选出的东北红豆杉B10细胞进行悬浮培养。结果发现,在诱导子的作用下,细胞内紫杉醇的含量在4d内大幅度提高,其中主要是促进了紫杉醇的两种前体化合物向紫杉醇的转化;细胞内高含量紫杉醇影响细胞的生长,可以采用两段培养法来提高紫杉醇的产量;同时,培养液中蔗糖、氮、磷、钾、镁等主要营养物质的消耗与紫杉醇的生物合成也存在密切的联系。
In order to use modern biotechnology to produce taxol, to study the biosynthesis of paclitaxel in cell suspension culture, the consumption of major nutrients in cell growth and culture fluids was studied. The inducer was added to Gamborg’s B5 medium and the suspension of B. cuspidata B10 cells induced by this laboratory was cultured. It was found that under the action of the elicitor, the content of paclitaxel in the cells was greatly increased within 4 days, which mainly promoted the conversion of paclitaxel’s two precursor compounds to paclitaxel; the high content of paclitaxel in the cells affected the growth of the cells. Two-stage culture was used to increase the production of taxol. At the same time, the consumption of major nutrients such as sucrose, nitrogen, phosphorus, potassium and magnesium in the culture fluid was also closely related to the biosynthesis of paclitaxel.