杜仲细胞悬浮培养产黄酮及其动力学研究

Study on Flavonoids Producing and Kinetics in Cell Suspension Culture of Eucommia ulmoides Oliv.

应用正交设计对杜仲细胞悬浮培养的基本培养基和植物生长物质浓度进行了筛选,并对影响杜仲细胞悬浮培养和总黄酮含量的不同因素进行了考察。结果表明,B5培养基+0.5mg/LNAA+0.6mg/L6-BA、蔗糖30g/L、初始pH5.0～5.5、接种量20g(FW)/L以及摇床转速110r/min为杜仲细胞悬浮培养的适宜条件。通过对杜仲悬浮细胞生长和代谢动力学的分析表明:杜仲细胞悬浮培养生长符合Logistic生长模型,最大比生长速率(μm)为0.417/d;细胞基于蔗糖的真正比生长得率(YG)与维持系数(m)分别为0.619g/g和0.0206g/(g.d-1);黄酮合成属部分生长耦联型,可用Luedeking-Piret模型进行描述。研究结果为杜仲细胞大规模悬浮培养生产天然活性成分奠定了基础。

The type of basic media and the contents of plant growth substances were investigated by orthogonal design experiment, and also the effects of different culture conditions on the growth of suspension cells and the accumulation of total flavonoids in Eucommia ulmoides were studied. The results showed that B5 medium supplemented with 0. 5mg/L NAA, 0.6mg/L 6-BA and 30g/L sucrose, at initial pH 5.0 -5.5, 20g （ FW）/L inoculation quantity and 110 r/min of rotation speed was a preferable culture conditions for E. ulmoides suspension cells growth and flavonoids synthesis. The results of metabolic kinetics analysis for E. ulmoides cell suspension culture showed that the logistic and Luedeking-Piret equations can be used for describing the kinetics of cell growth, sucrose consumption and flavonoids production during the process. The maximum specific growth rate （μm）, the actual growth yield based on sucrose （Yc） and maintenance coefficient （m） were 0. 417/d, 0. 619g/g and 0. 0206g/（ g · d^-1） respectively. All these outcomes could give a basis for establishing the suspension cell culture of E. ulmoides and production of the natural active components in large-scale.