Rapid chromatographic method for the isolation of glucose-6-phosphate dehydrogenase from yeast enzyme concentrate

A simple method for the isolation of glucose-6-phosphate dehydrogenase (G6PDH) from yeast enzyme concentrate is described. The method is based on high-performance anion-exchange chromatography and was developed in three steps: (1) optimization of chromatographic conditions on Polyanion SI-8 microns, (2) transfer of the optimized conditions to a preparative column containing Polyanion SI-17 microns and (3) final purification on an analytical column. As the enzyme is a minor component of the crude mixture, its presence is detected by a simple enzymatic method. The purity of the final fraction is checked by polyacrylamide gel electrophoresis.