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

Abstract

The influence of sugars in pre-cultivation media suspended plant cells on the kinetics of the sugar uptake and the ethanol production was studied by mid-infrared spectroscopy using a Fourier transform infrared spectrometer (FT-IR) equipped with an attenuate total reflection accessory (ATR). We performed the plant cell cultivation with Nicotiana tabacum cv. Bright Yellow No.2 (TBY-2) cells and Oryza sativa L., Japonica, cv. Nipponbare (rice) cells, respectively, in pre-culture and culture media, which had various types of glucose, fructose, sucrose or glucose-fructose mixtures. The results confirmed the kinetic differences between the TBY-2 cells and rice cells. These results suggested that the TBY-2 cells consumed sugar before growth and the rice cells consumed sugar after growth, moreover, the ethanol content increased just after cell growth was activated based on the non-dimensional cultivation time for the cell growth behavior.

Analysis of kinetic uptake phenomena of monosaccharide and disaccharide by suspension TBY-2 cells using an FT-IR/ATR method

“…This indicated that a part or most of the trehalose in the medium was hydrolyzed to glucose, which was consumed by the TBY-2 cell. Moreover, in the sucrose medium, the sucrose concentration decreased with an increase in the glucose and fructose concentrations, which the sucrose in the medium was hydrolyzed similar to the previous studies [13][14][15][16]32]. In contrast to these results, in the maltose medium, we rarely observed any sugar other than maltose by both the FT-IR/ATR and HPLC method (data not shown).…”

“…The ATR spectra were obtained with a horizontal zinc selenide ATR sampling accessory (GRASEBY SPE-CAC, SPECACLAMP ATR 11080). The dry cell weight after drying was compared to the turbidity of culture medium by at 600 nm as described previously [13][14][15][16]28].…”

“…Furthermore, we studied the specific uptake rates of sugar, which is one of the most important indices characterizing the sugar metabolic kinetics of cells. We then determined the specific uptake rates after quantification of the sugar uptake rate and the cell density according to the method provided in our paper [14,16,28] as follows.…”

“…Bright Yellow No.2 (TBY-2), which were sub-cultured in the Murashige-Skoog (MS) [27] medium for 7 days at 298 K on a rotary shaker (150 rpm) in the dark as described in the previous paper [13][14][15][16]28].…”

“…Using the FT-IR/ATR method developed by Hashimoto et al [15], we studied the influence of sugar species in the pre-culture and culture media on the kinetic sugar uptake phenomena and the ethanol production by comparing the characteristics of the TBY-2 cells with the rice cells [16].…”