Hashimoto A, Nakanishi K, Motonaga Y, Kameoka T
Department of Sustainable Resource Sciences, Faculty of Bioresources, Mie University, Tsu, Mie 514-8507, Japan.
Biotechnol Prog. 2001 May-Jun;17(3):560-4. doi: 10.1021/bp010013w.
A simple, rapid and accurate evaluation of the sugar uptake rate of suspended plant cells from culture media was developed with the predicted sugar contents measured by mid-infrared spectroscopy using a Fourier transform infrared (FT-IR) spectrometer equipped with an attenuated total reflectance (ATR) accessory. We performed plant cell cultivation with Nicotiana tabacum cv. Bright Yellow No.2 (TBY-2) in culture media, which had various combinations of glucose, fructose and sucrose concentrations at the initial stage, and measured simultaneously each sugar content in the medium by the FT-IR/ATR method. By applying a logistic function to the predicted sugar contents and cell density in the medium during cultivation, the specific sugar uptake rates by the suspended TBY-2 cells were easily and continuously obtained. Thus the kinetic sugar uptake phenomena by the TBY-2 cells were well confirmed overall using the developed method. Additionally it was found that the fraction of sucrose of the initial total sugar content might kinetically affect the sugar uptake process and cell growth. Also, the relationship between the nondimensional cell density and sucrose content could be classified into three groups on the basis of the initial fraction of sucrose.
利用配备衰减全反射(ATR)附件的傅里叶变换红外(FT-IR)光谱仪通过中红外光谱法测量预测的糖含量,开发了一种简单、快速且准确的评估悬浮植物细胞从培养基中摄取糖速率的方法。我们使用烟草品种亮黄2号(TBY-2)在培养基中进行植物细胞培养,培养基在初始阶段具有葡萄糖、果糖和蔗糖浓度的各种组合,并通过FT-IR/ATR方法同时测量培养基中的每种糖含量。通过将逻辑函数应用于培养过程中培养基中预测的糖含量和细胞密度,可轻松且连续地获得悬浮的TBY-2细胞的比糖摄取速率。因此,使用所开发的方法总体上很好地证实了TBY-2细胞的动力学糖摄取现象。此外,还发现初始总糖含量中蔗糖的比例可能在动力学上影响糖摄取过程和细胞生长。而且,基于蔗糖的初始比例,无量纲细胞密度与蔗糖含量之间的关系可分为三组。