Chemometric analysis of the oscillatory kinetics of glycolytic intermediates in a yeast extract from data measured by FTIR-spectroscopy

Petro Khoroshyy1, László Zimányi1, Thomas Mair2, Andrea Müller2, and Stefan C. Müller2

1Institute of Biophysics, Biological Research Center of the Hungarian Academy of Sciences, Szeged, Hungary, and 2Otto-von-Guericke-Universitaet, Institute of Experimental Physics, Group of Biophysics, Magdeburg, Germany

In the present work we demonstrate that FTIR-spectroscopy is a powerful tool for the time resolved and noninvasive measurement of multi substrate/product interactions in complex metabolic networks as exemplified by the oscillating glycolysis in a yeast extract. We found that many of the glycolytic intermediates can be identified with FTIR-spectroscopy. For this, we have constructed a spectral library of most of the glycolytic intermediates and obtained the kinetics of single components in spectra from glycolysing yeast extract by the use of mathematical fitting procedures. We compare the results obtained by direct fit of the yeast extract spectra with the spectra of the pure chemical components to the results provided by more advanced chemometric methods. In particular, singular value decompostition and independent component analysis of the spectrotemporal data matrix were performed. The results are in good agreement with the known phase relationships of oscillatory glycolysis. They provide the basis for future application of this method to investigate the energy metabolism of living cells.

Email: petro@nucleus.szbk.u-szeged.hu

Address: Institute of Biophysics, Biological Research Center of the Hungarian Academy of Sciences, Temesvari krt. 62., P.O.Box 521, Szeged, Hungary, H-6701,