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Literature summary for 1.1.99.37 extracted from

  • Hektor, H.J.; Kloosterman, H.; Dijkhuizen, L.
    Identification of a magnesium-dependent NAD(P)(H)-binding domain in the nicotinoprotein methanol dehydrogenase from Bacillus methanolicus (2002), J. Biol. Chem., 277, 46966-46973.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Bacillus methanolicus

Protein Variants

Protein Variants Comment Organism
D100N lacks most of the bound cofactor NAD(H) and has lost all coenzyme NAD+-dependent MDH activity Bacillus methanolicus
D88N only minor differences to wild-type in kinetics Bacillus methanolicus
G15A only minor differences to wild-type in kinetics Bacillus methanolicus
G95A impaired in cofactor NAD(H) binding but retains coenzyme NAD+-dependent MDH activity Bacillus methanolicus
G95A low MDH activity, 10fold decrease in affinity for NAD+/NADH, insensitive to activator protein Bacillus methanolicus
K103R lacks most of the bound cofactor NAD(H) and has lost all coenzyme NAD+-dependent MDH activity Bacillus methanolicus
S97G impaired in cofactor NAD(H) binding but retains coenzyme NAD+-dependent MDH activity Bacillus methanolicus
S97G insensitive to activator protein, but displays fully activated MDH reaction rates Bacillus methanolicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Km value of wild-type for NAD+ 0.04 mM, for NADH 0.01 mM, in absence of Mg2+, and for NAD+ 0.03 mM, for NADH 0.011 mM, in presence of Mg2+, of respectively Bacillus methanolicus

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
Bacillus methanolicus
Zn2+
-
Bacillus methanolicus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
43000
-
10 * 43000, SDS-PAGE Bacillus methanolicus

Organism

Organism UniProt Comment Textmining
Bacillus methanolicus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Bacillus methanolicus

Reaction

Reaction Comment Organism Reaction ID
methanol + acceptor = formaldehyde + reduced acceptor Ping-Pong type of reaction mechanism Bacillus methanolicus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.413
-
pH 9.5, 50°C Bacillus methanolicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
-
Bacillus methanolicus methanol + N,N-dimethyl-4-nitrosoaniline
-
r
methanol + N,N-dimethyl-4-nitrosoaniline
-
Bacillus methanolicus formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
-
r

Subunits

Subunits Comment Organism
decamer 10 * 43000, SDS-PAGE Bacillus methanolicus

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Bacillus methanolicus
NADH
-
Bacillus methanolicus