Theoretical work has suggested that a cell-free process, consisting of the twelve enzymes involved in the yeast anaerobic glycolysis pathway, is capable of making ethanol much faster than a microorganism based fermentation.  This and other potential advantages have suggested that a cell-free process may improve the economy of fuel ethanol production.  We have attempted to further investigate the potential of this process with lab scale studies.  Glycolytic enzymes were purified from yeast and a micro scale assay was developed to evaluate varying enzyme, substrate and cofactor concentrations.  Progress of the reactions was followed by monitoring ethanol produced as well as observing phosphorylated metabolite profiles using ³¹P NMR.  Correlation of the results to theoretical predictions will be detailed.