Electron microscopic analysis and biochemical characterization of a novel methanol dehydrogenase from the thermotolerant Bacillus sp. C1.
Methanol dehydrogenase from the thermotolerant Bacillus sp. C1 was studied by electron microscopy and image processing. Two main projections can be distinguished: one exhibits 5-fold symmetry and has a diameter of 15 nm, the other is rectangular with sides of 15 and 9 nm. Subsequent image processing showed that the 5-fold view possesses mirror symmetry. The rectangular views can be divided into two separate classes, one of which has 2-fold rotational symmetry. It is concluded that methanol dehydrogenase is a decameric molecule, and a tentative model is presented. The estimated molecular weight is 430,000, based on a subunit molecular weight of 43,000. The enzyme contains one zinc and one to two magnesium ions per subunit. N-terminal amino acid sequence analysis revealed substantial similarity with alcohol dehydrogenases from Saccharomyces cerevisiae, Zymomonas mobilis, Clostridium acetobutylicum, and Escherichia coli, which contain iron or zinc but no magnesium. In view of the aberrant structural and kinetic properties, it is proposed to distinguish the enzyme from common alcohol dehydrogenases (EC 1.1.1.1) by using the name NAD-dependent methanol dehydrogenase.
Vonck
J
J
Biochemical Laboratory, University of Groningen, The Netherlands.
Arfman
N
N
De Vries
G E
GE
Van Beeumen
J
J
Van Bruggen
E F
EF
Dijkhuizen
L
L
D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000648:ultrastructure; D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology*; D006358:Hot Temperature; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012689:Sequence Homology, Nucleic Acid
D000429:Alcohol Oxidoreductases; C019641:alcohol dehydrogenase (acceptor)