三尖杉悬浮细胞的培养及抗癌生物碱的产生

建立三尖杉悬浮细胞培养系统（ＭＳ＋ＮＡＡ ３ ｍｇ／Ｌ＋ＫＴ ０．１ｍｇ／Ｌ＋庶糖 ３％）优化培养条件，产生更多的抗癌生物碱。方法：研究不同环境因子：接种量、摇床转速、激素、氮源、碳源、前体及诱导子调控对三尖杉悬浮细胞的生长及三尖杉酯碱和高三尖杉酯碱产生的影响；以及细胞生长和产物积累的偶联反应。结果：三尖杉悬浮细胞的生长与三尖杉酯类碱的产生呈正相关；接种量、激素与Ｄ－剂（生长素）组合；碳源；前体苯丙氨酸有利于细胞生长及酯类碱的产生。摇床转速；氮源利于细胞生长。激素与ＢＲ（油菜内酯素）的组合；无机氮ＮＯ_３－；蛋白胨；诱导子能显著提高三尖杉酯类碱的含量。结论：择优筛选出有利于三尖杉酯类生物碱的生物、化学、物理因子，是提高悬浮细胞抗癌生物碱的有效途径。

Production of Antitumor Alkaloids from Suspension Cell Culture of Cephalotaxus Fortunei

Purpose: The suspension cell culture system of Cephalotaxus fortunei Hook. f (MS NAA 3 mg/L KT 0.1 mg/L sucrose 3% ) was established in this original research. Methods: The effects of different environmental factors, inoculation amount, shaking sped, auxin, elicitor, precursor, nitrogen resource and carbon resource on the production of harringtonine or homoharringtonine were studied. The relationship between biomass and product synthesis was also studied. Resulta: The results showed that higher inoculation amount was helpful for the cell growth and product synthesis. The condition: inoculation mass of 2g/ml, auxin combined with D-reagent, 1.5% sucrose and 1. 5% glucose,phenylalanine as the precursor not only was optimum for cell growth but also could increase the cell dry weight for the homoharringtonine synthesis. The shaking sped of 80r/min and beef extract broth as nitrogen resources was the best for cell growth. Auxin combined with BR, inorganic nitrogen in NO_3-and peptone gel plus clicitor can significantly increase harringtonine production. Conclusion: Therefore, choosing the optimum biological, chemical and physical conditions is an efficient way to increase the production of harringtonine.