The application of triazine dye affinity chromatography to the large-scale purification of glycerokinase from Bacillus stearothermophilus.
Gram quantities of homogeneous glycerokinase have been prepared from the thermophilic bacterium, Bacillus stearothermophilus, using three major steps: precipitation of debris at pH 5.1, ion-exchange chromatography on DEAE-Sephadex, and affinity chromatography on Procion Blue MX-3G-Sepharose. This method is a considerable improvement over conventional techniques; the purified enzyme was obtained with a 40% recovery and a specific activity of 120 units (mumol/min)/mg protein. A modified culture medium enabled yields of 3.4 X 10(6) units of enzyme to be obtained from 400-liter production cultures.
D002846:Chromatography, Affinity / Q000379:methods; D002852:Chromatography, Ion Exchange; D004396:Coloring Agents; D003470:Culture Media; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D005991:Glycerol Kinase / Q000302:isolation & purification*; D010770:Phosphotransferases / Q000302:isolation & purification*
D004396:Coloring Agents; D003470:Culture Media; D010770:Phosphotransferases; D005991:Glycerol Kinase