An improved method for the purification of Bacillus subtilis glucose dehydrogenase cloned in Escherichia coli.
An improved and simplified purification procedure has been developed for the isolation of the Bacillus subtilis glucose dehydrogenase which has resulted in a 10 fold higher yield of pure enzyme. The purification procedure utilizes gene cloning and an additional ammonium sulfate step to facilitate the removal of contaminating proteins. The procedure requires fewer chromatographic steps than previously reported, thus simplifying the procedure. This improved and simplified purification of B. subtilis glucose dehydrogenase will facilitate further structure-function studies of this sporulation specific enzyme.
D001412:Bacillus subtilis / Q000201:enzymology*; D002237:Carbohydrate Dehydrogenases / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D042843:Glucose 1-Dehydrogenase; D005948:Glucose Dehydrogenases / Q000302:isolation & purification*; D010957:Plasmids
D002237:Carbohydrate Dehydrogenases; D005948:Glucose Dehydrogenases; D042843:Glucose 1-Dehydrogenase