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

A simple, rapid and accurate mid-infrared (MIR) spectroscopic method for simultaneously determining the product (ethanol) content and the nutrient (sugar) content in plant-cell culture media was developed using a Fourier transform infrared (FT-IR) spectrometer equipped with an attenuated total reflectance (ATR) accessory. We assessed the potential of this method by comparing it to a high-performance liquid chromatography (HPLC) method, and using the developed method to measure the ethanol and sugar contents simultaneously in liquid culture media with rice and tabacum cell suspensions, respectively. The experimental results suggest that the sugar consumption and ethanol production behaviors of the plant cell suspensions can be non-destructively and simultaneously monitored using the developed method. Furthermore, the spectroscopic method provided in this study could be developed into a technique that could be used to analyze the overall kinetics of the metabolism of the plant cell suspensions.

[1]  Jack L. Koenig,et al.  Infrared and raman spectroscopy of carbohydrates : Part IV. Identification of configuration- and conformation-sensitive modes for D-glucose by normal coordinate analysis , 1974 .

[2]  P. Bernath,et al.  Spectra of Atoms and Molecules , 1996 .

[3]  B. McNeil,et al.  Near-Infrared Spectroscopic Determination of Acetate, Ammonium, Biomass, and Glycerol in an Industrial Escherichia Coli Fermentation , 1996 .

[4]  E. K. Kemsley,et al.  Detection of adulteration of raspberry purees using infrared spectroscopy and chemometrics , 1996 .

[5]  S. Sivakesava,et al.  Simultaneous determination of multiple components in lactic acid fermentation using FT-MIR, NIR, and FT-Raman spectroscopic techniques , 2001 .

[6]  K. Lindsey Plant Tissue Culture Manual , 1992, Springer US.

[7]  Atsushi Hashimoto,et al.  Mid-Infrared Spectroscopic Measurement of Ionic Dissociative Materials in the Metabolic Pathway , 2003, Applied spectroscopy.

[8]  篤 橋本,et al.  FT-IR/ATR法を用いた糖水溶液の赤外分光分析 , 1998 .

[9]  Quantitative analysis by mid-infrared spectrometry in food and agro-industrial fields , 1993 .

[10]  T. Kameoka,et al.  Effects on pre-cultivation on sugar metabolic kinetics of suspension plant cells measured by mid-infrared spectroscopy , 2003 .

[11]  Sugar Metabolic Analysis of Suspensions of Plant Cells Using an FT‐IR/ATR Method , 2001, Biotechnology progress.

[12]  E. Schaller,et al.  Analytical methods for the determination of the geographic origin of Emmental cheese: mid- and near-infrared spectroscopy , 2003 .

[13]  J. Sastre Toraño,et al.  Quantitative analysis of active compounds in pharmaceutical preparations by use of attenuated total-reflection Fourier transform mid-infrared spectrophotometry and the internal standard method , 2001, Fresenius' journal of analytical chemistry.

[14]  F. Skoog,et al.  A revised medium for rapid growth and bio assays with tobacco tissue cultures , 1962 .

[15]  S. Schmidt,et al.  Near infrared spectroscopy in fermentation and quality control for amino acid production , 1998 .

[16]  K. Shimamoto,et al.  Transformation and regeneration of rice protoplasts , 1991 .

[17]  T. Kameoka,et al.  Mid-Infrared Spectroscopic Determination of Sugar Contents in Plant-Cell Culture Media Using an ATR Method , 2000 .

[18]  M. Saigusa,et al.  Studies on the nutrition of rice cell culture II. Microelement requirement and the effects of deficiency , 1975 .

[19]  P. Salmon,et al.  Real-time analyte monitoring of a fungal fermentation, at pilot scale, using in situ mid-infrared spectroscopy , 2001 .

[20]  T. Pan,et al.  Development of a quantification system of ionic dissociative metabolites using an FT-IR/ATR method , 2003, Bioprocess and biosystems engineering.

[21]  篤 橋本,et al.  食品糖成分分析へのFT-IR/ATR法の応用 , 1998 .

[22]  N. J. Harrick,et al.  Internal Reflection Spectroscopy Review and Supplement , 1985 .

[23]  Bernhard Lendl,et al.  Simultaneous determination of enzyme activities by FTIR-spectroscopy in an one-step assay , 1999 .

[24]  F. Skoog,et al.  A revised medium for the growth and bioassay with tobacco tissue culture , 1962 .

[25]  N. Harrick,et al.  Internal reflection spectroscopy , 1968 .

[26]  V. Bellon-Maurel,et al.  Quantitative Analysis of Individual Sugars during Starch Hydrolysis by FT-IR/ATR Spectrometry. Part I: Multivariate Calibration Study—Repeatibility and Reproducibility , 1995 .

[27]  K. Ohira,et al.  Studies on the nutrition of rice cell culture I. A simple, defined medium for rapid growth in suspension culture : , 1973 .

[28]  Mark A. Arnold,et al.  Simultaneous Measurements of Glucose, Glutamine, Ammonia, Lactate, and Glutamate in Aqueous Solutions by Near-Infrared Spectroscopy , 1996 .

[29]  J. Bosset,et al.  Analytical methods for the determination of the geographic origin of Emmentaler cheese. Main framework of the project; chemical, biochemical, microbiological, colour and sensory analyses , 2002 .

[30]  J. Koenig,et al.  Infrared and raman spectroscopy of carbohydrates , 1971 .

[31]  S. Arnold,et al.  Attenuated total reflectance Fourier transform mid-infrared spectroscopic quantification of sorbitol and sorbose during a Gluconobacter biotransformation process , 2003, Biotechnology Letters.

[32]  Y. Nakano,et al.  Determination of the Content of Water and Rice Bran in Solid Media Used for Mushroom Cultivation Using Near-Infrared Spectroscopy , 1998 .