Quinoprotein D-glucose dehydrogenases in Acinetobacter calcoaceticus LMD 79.41: purification and characterization of the membrane-bound enzyme distinct from the soluble enzyme.
Acinetobacter calcoaceticus is known to contain soluble and membrane-bound quinoprotein D-glucose dehydrogenases while other oxidative bacteria such as Pseudomonas or Gluconobacter contain only membrane-bound enzyme. The two different forms were believed to be the same enzyme or interconvertible. Present results show that the two different forms of glucose dehydrogenase are distinct from each other in their enzymatic and immunological properties as well as in their molecular size. The soluble and membrane-bound glucose dehydrogenases were separated after French press-disruption by repeated ultracentrifugation, and then purified to nearly homogeneous state. The soluble enzyme was a polypeptide of 55 Kdaltons, while the membrane-bound enzyme was a polypeptide of 83 Kdaltons which is mainly monomeric in detergent solution. Both enzymes showed different enzymatic properties including substrate specificity, optimum pH, kinetics for glucose, and reactivity for ubiquinone-homologues. Furthermore, the two enzymes could be distinguished immunochemically; the membrane-bound enzyme is cross-reactive with an antibody raised against membrane-bound enzyme purified from Pseudomonas but not with antibody elicited against the soluble enzyme, while the soluble enzyme is not cross-reactive with the antibody of membrane-bound enzyme. Data also suggest that the membrane-bound enzyme functions by linking to the respiratory chain via ubiquinone though the function of the soluble enzyme remains unclear.
Matsushita
K
K
Department of Agricultural Chemistry, Faculty of Agriculture, Yamaguchi University, Japan.
Shinagawa
E
E
Adachi
O
O
Ameyama
M
M
D000150:Acinetobacter / Q000201:enzymology*; D002237:Carbohydrate Dehydrogenases / Q000302:isolation & purification* / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D003429:Cross Reactions; D005948:Glucose Dehydrogenases / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008565:Membrane Proteins / Q000276:immunology / Q000378:metabolism*; D008970:Molecular Weight; D045542:PQQ Cofactor; D015363:Quinolones / Q000378:metabolism; D012995:Solubility; D014451:Ubiquinone / Q000031:analogs & derivatives / Q000378:metabolism
D008565:Membrane Proteins; D015363:Quinolones; D014451:Ubiquinone; D045542:PQQ Cofactor; D002237:Carbohydrate Dehydrogenases; D005948:Glucose Dehydrogenases; C033955:glucose dehydrogenase (pyrroloquinoline-quinone)