PMID:9003445
| Citation |
Hanschmann, H and Kleber, HP (1997) Purification and characterization of D(+)-carnitine dehydrogenase from Agrobacterium sp.--a new enzyme of carnitine metabolism. Biochim. Biophys. Acta 1337:133-42 |
|---|---|
| Abstract |
D(+)-Carnitine dehydrogenase from Agrobacterium sp. catalyzes the oxidation of D(+)-carnitine to 3-dehydrocarnitine as initial step of D(+)-carnitine degradation. The NAD(+)-specific, cytosolic enzyme was purified 126-fold to apparent electrophoretic homogeneity by 4 chromatographic steps. The molecular mass of the native enzyme was estimated to be 88 kDa by size-exclusion chromatography. It seems to be composed of 3 identical subunits with a relative molecular mass of 28 kDa as found by sodium dodecyl sulfate polyacrylamide gel electrophoresis and laser-induced mass spectrometry. The isoelectric point was found to be 4.7-5.0. The optimum temperature is 37 degrees C and the optimum pH for the oxidation and the reduction reaction are 9.0-9.5 and 5.5-6.5, respectively. The purified enzyme was further characterized with respect to substrate specificity, kinetic parameters and amino terminal sequence. Analogues of D(+)-carnitine (L(-)-carnitine, crotonobetaine, gamma-butyrobetaine, carnitine amide, glycine betaine, choline) are competitive inhibitors of D(+)-carnitine oxidation. The equilibrium constant of the reaction of D(+)-carnitine dehydrogenase was determined to be 2.2 x 10(-12). The purified D(+)-carnitine dehydrogenase has similar kinetic properties to the L(-)-carnitine dehydrogenase from the same microorganism as well as to L(-)-carnitine dehydrogenases of other bacteria. |
| Links | |
| Keywords |
Alcohol Oxidoreductases/antagonists & inhibitors; Alcohol Oxidoreductases/isolation & purification; Alcohol Oxidoreductases/metabolism; Amino Acid Sequence; Bacterial Proteins/isolation & purification; Bacterial Proteins/metabolism; Carnitine/analogs & derivatives; Carnitine/metabolism; Chromatography/methods; Cytosol/enzymology; Enzyme Inhibitors; Isoelectric Point; Isomerism; Kinetics; Molecular Sequence Data; Molecular Weight; NAD/metabolism; Rhizobium/enzymology; Sequence Analysis; Sequence Homology, Amino Acid; Substrate Specificity |
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