Purification and properties of membrane-bound D-sorbitol dehydrogenase from Gluconobacter suboxydans IFO 3255.
D-Sorbitol dehydrogenase was solubilized from the membrane fraction of Gluconobacter suboxydans IFO 3255 with Triton X-100 in the presence of D-sorbitol. Purification of the enzyme was done by fractionation with column chromatographies of DEAE-Cellulose, DEAE-Sepharose, hydroxylapatite, and Sephacryl HR300 in the presence of Triton X-100. The molecular mass of the enzyme was 800 kDa, consisting of homologous subunits of 80 kDa. The optimum pH of the enzyme activity was 6.0, and the optimum temperature was 30 degrees C. Western blot analysis suggested the occurrence of the enzyme in all the Gluconobacter strains tested.
Sugisawa
Teruhide
T
Department of Applied Microbiology, Nippon Roche Research Center, Kamakura, Kanagawa, Japan. teruhide.sugisawa@roche.com
Hoshino
Tatsuo
T
D019342:Acetic Acid; D001672:Biocompatible Materials; D002384:Catalysis; D002462:Cell Membrane / Q000201:enzymology; D002848:Chromatography, DEAE-Cellulose / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004791:Enzyme Inhibitors; D004795:Enzyme Stability; D020576:Gluconobacter / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D019807:Iodoacetic Acid / Q000494:pharmacology; D007064:L-Iditol 2-Dehydrogenase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008670:Metals; D011487:Protein Conformation; D011803:Quinine / Q000494:pharmacology; D012967:Sodium Dodecyl Sulfate; D012995:Solubility; D013012:Sorbitol / Q000378:metabolism; D013013:Sorbose / Q000378:metabolism; D013379:Substrate Specificity; D013696:Temperature
D001672:Biocompatible Materials; D004791:Enzyme Inhibitors; D008670:Metals; D012967:Sodium Dodecyl Sulfate; D013012:Sorbitol; D017886:Durapatite; D011803:Quinine; D007064:L-Iditol 2-Dehydrogenase; D013013:Sorbose; D019342:Acetic Acid; D019807:Iodoacetic Acid