A simple procedure to obtain yeast hexokinase free of glucosephosphate isomerase and mannosephosphate isomerase.
A hexokinase preparation was obtained from a Saccharomyces cerevisiae mutant strain deficient in glucosephosphate isomerase (GPI) and mannosephosphate isomerase (MPI) by precipitation with ammonium sulfate. The supernatant fraction corresponding to 40-60 % saturation showed the lowest content in GPI and MPI activity. The fraction was used without further purification in the determination of glucose, either free or in a mixture with fructose and mannose. The results were similar to those obtained with pure commercial hexokinase.
D000645:Ammonium Sulfate; D011232:Chemical Precipitation; D005956:Glucose-6-Phosphate Isomerase / Q000235:genetics; D006593:Hexokinase / Q000302:isolation & purification*; D008359:Mannose-6-Phosphate Isomerase / Q000235:genetics; D009154:Mutation; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics
D006593:Hexokinase; D008359:Mannose-6-Phosphate Isomerase; D005956:Glucose-6-Phosphate Isomerase; D000645:Ammonium Sulfate