VIEW ARTICLE    DOI: 10.1094/ASBCJ-41-0125

Glycolytic Flux in Lager Yeast Fermentations: Application of Isotachophoresis and Bioluminescence for Measurement of Cellular Intermediates. D. S. Ryder, D. R. Woods, M. Castiau, and C. A. Masschelein. J. Am. Soc. Brew. Chem. 41:0125, 1983.

The relationship between yeast growth and glycolytic efficiency per unit of yeast during fermentation was examined at a fundamental level. The study required that intracellular nucleotides, organic acids, and hexose phosphates be recovered in solution. Measurement techniques using the concepts of isotachophoresis and bioluminescence were developed to achieve desired specificity and sensitivity (picomole range). Cell permeabilization and metabolite extraction were achieved using a procedure mediated by boiling water. In spite of its simplicity, this procedure appeared more efficient than alternative methods on the basis of recovering higher amounts of nucleotides. Isotachophoresis was used for the quantitative measurement of organic acids, and adenosine, guanosine, uridine, and cytidine, mono-, di-, and triphosphates. To separate nucleotides, discrete intermediate-mobility "spacers" were added to the sample solution, and steady-state mixing techniques were applied. Quantitative measurement of glucose-6-phosphate, fructose-6-phosphate, fructose-1,6-bisphosphate, and triose phosphate was achieved by coupled enzymatic assays and a Photobacterial bioluminescence system. The techniques described are playing a useful role in explaining the regulation of glycolytic flux in lager yeast fermentations.

Keywords: Bioluminescence, Hexose phosphates, Isotachophoresis, Nucleotides, Organic acids, Permeabilized cells