Enhanced glucose to fructose conversion in acetone with xylose isomerase stabilized by crystallization and cross-linking

The effects of acetone and ethanol on glucose to fructose conversion catalyzed by soluble and cross-linked crystalline (CLXIC) xylose isomerase were studied. Relative to pure buffer solvent, the fructose production rate was more than doubled in 50% acetone. The same kind of increase in the isomerization rate was not seen with ethanol. Increase both in acetone and in ethanol concentration in the reaction solvent enhanced the production of fructose. At 50 degrees C in pure buffer solvent the reaction mixture contained 49% fructose in equilibrium and in 90% acetone the fructose equilibrium content was 64%. Furthermore, CLXIC was relatively stable in the presence of high concentration of acetone: 70-80% of activity was left after incubation for 24 h at 50 degrees C in buffer solutions (pH 7.2) containing 10-90% acetone. In buffer containing 50% ethanol only 2% of the initial activity of CLXIC was retained after 24 h at 50 degrees C. Soluble xylose isomerase was considerably less stable than CLXIC in both acetone- and ethanol-containing solutions. These results show that the addition of acetone enhances the production of fructose from glucose by enhancing the reaction rate and shifting the equilibrium toward fructose. However, xylose isomerase must be in the form of cross-linked crystals for maximal activity and stability.