Sugar Metabolic Analysis of Suspensions of Plant Cells Using an FT‐IR/ATR Method

Abstract

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.

MIR spectroscopic analysis on sugar metabolic and ethanol productive kinetics of suspension TBY-2 and rice cells pre-cultured in various media

“…The ATR spectra were obtained with a horizontal zinc selenide ATR sampling accessory (GRASEBY SPECAC, SPECACLAMP ATR 11080). The dry cell weight was compared to the turbidity of the culture medium using a UV-VIS-NIR scanning spectrophotometer as previously described [14,15,18,19].…”

“…This study was carried out with the TBY-2 and rice cells, which were essentially sub-cultured under the same condition as that described in the previous paper [14,15,18,19].…”

Simple and rapid determination of metabolite content in plant cell culture medium using an FT-IR/ATR method

“…In our previous study [20], we developed a method of determining the sugar content in MS media using FT-IR/ATR, that focused on sugar uptake by TBY-2 cells [21]. In this study, we investigated the simultaneous quantification of both the nutrient sugars and the product ethanol.…”

“…As a result, the content of each sugar in the culture medium in which the plant cells were suspended was simultaneously and accurately measured by MIR spectroscopy. Additionally, a simple, rapid and accurate method for analyzing sugar metabolic kinetics by the FT-IR/ATR method was developed [21][22][23]. Also, the FT-IR/ATR method has the potential to theoretically determine most of the other chemical components relating to the metabolism besides the sugars.…”

Development of a quantification system of ionic dissociative metabolites using an FT-IR/ATR method

“…The accuracy of the developed method was almost comparable to that of a high-performance liquid chromatography method [5]. The kinetic sugar uptake phenomena by suspending cells were overall well confirmed using the developed method [7].…”