title: [Purification and characteristics of methanol dehydrogenase of Methylobacterium nodulans rhizosphere phytosymbionts].
abstract: Methanol dehydrogenase (MDG) of the facultative methylotrophic phytosymbiont Methylobacterium nodulans has been purified for the first time to an electrophoretically homogeneous state and characterized. The native protein with a molecular mass of 70 kDa consists of large (60 kDa) and small (6 kDa) subunits. The purified protein displayed a specter identical to that of pyrochinolinchinon (PCC)-containing MDGs (pI 8.7, pH optimum in the range 9-10). The enzyme was inactive in the absence of ammonium or methylamine and exhibited a wide substrate specificity with regard to C1-C2 alcohols with the highest affinity to methanol (K(M) = 70 mM), but it did not oxidize benzyl and secondary alcohols. The apparent values of K(M) to primary alcohols increased with the length of the carbonic chain. The enzyme was characterized by a high stability level even in the absence of a substrate. An immobilized enzyme was used for amperometric methanol detection.
authors: T A
authors: TA
authors: Kuznetsova
authors: A P
authors: AP
authors: Beschastnyĭ
authors: O N
authors: ON
authors: Ponomareva
authors: Iu A
authors: IuA
authors: Trotsenko
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification*; D000641:Ammonia / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D015374:Biosensing Techniques; D029864:Crotalaria / Q000382:microbiology; D055664:Electrochemical Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D000432:Methanol / Q000032:analysis* / Q000737:chemistry; D008744:Methylamines / Q000737:chemistry; D020580:Methylobacterium / Q000737:chemistry / Q000201:enzymology*; D008970:Molecular Weight; D021122:Protein Subunits / Q000737:chemistry / Q000302:isolation & purification*; D058441:Rhizosphere; D013379:Substrate Specificity; D013559:Symbiosis
chemical_list: D001426:Bacterial Proteins; D004800:Enzymes, Immobilized; D008744:Methylamines; D021122:Protein Subunits; D000641:Ammonia; C027451:methylamine; D000429:Alcohol Oxidoreductases; C019641:alcohol dehydrogenase (acceptor); D000432:Methanol