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Literature summary 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

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.1.1.244 additional information strongly stimulated by the ACT protein Bacillus methanolicus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.244 expressed in Escherichia coli DH5alpha Bacillus methanolicus
1.1.99.37 expression in Escherichia coli Bacillus methanolicus

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.244 D100N strongly reduced NAD+-binding, no activity Bacillus methanolicus
1.1.1.244 D88N only minor effects on activity Bacillus methanolicus
1.1.1.244 G13A only minor effects on activity Bacillus methanolicus
1.1.1.244 G15A only minor effects on activity Bacillus methanolicus
1.1.1.244 G95A impaired cofactor binding, low acitivity Bacillus methanolicus
1.1.1.244 K103R strongly reduced NAD+-binding, no activity Bacillus methanolicus
1.1.1.244 S97G impaired cofactor binding, much higher acitivity than the wild type enzyme, no activation with ACT protein Bacillus methanolicus
1.1.1.244 S97T impaired cofactor binding, much higher acitivity than the wild type enzyme, no activation with ACT protein Bacillus methanolicus
1.1.99.37 D100N lacks most of the bound cofactor NAD(H) and has lost all coenzyme NAD+-dependent MDH activity Bacillus methanolicus
1.1.99.37 D88N only minor differences to wild-type in kinetics Bacillus methanolicus
1.1.99.37 G15A only minor differences to wild-type in kinetics Bacillus methanolicus
1.1.99.37 G95A impaired in cofactor NAD(H) binding but retains coenzyme NAD+-dependent MDH activity Bacillus methanolicus
1.1.99.37 G95A low MDH activity, 10fold decrease in affinity for NAD+/NADH, insensitive to activator protein Bacillus methanolicus
1.1.99.37 K103R lacks most of the bound cofactor NAD(H) and has lost all coenzyme NAD+-dependent MDH activity Bacillus methanolicus
1.1.99.37 S97G impaired in cofactor NAD(H) binding but retains coenzyme NAD+-dependent MDH activity Bacillus methanolicus
1.1.99.37 S97G insensitive to activator protein, but displays fully activated MDH reaction rates Bacillus methanolicus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.244 0.02
-
 NAD+ native enzyme, pH 9.5, 50°C Bacillus methanolicus
1.1.1.244 0.04
-
 NAD+ recombinant enzyme without added Mg2+, pH 9.5, 50°C Bacillus methanolicus
1.1.1.244 0.2
-
 NAD+ S97T mutant enzyme without added Mg2+, pH 9.5, 50°C Bacillus methanolicus
1.1.1.244 2.5
-
 NAD+ S97G mutant enzyme without added Mg2+, pH 9.5, 50°C Bacillus methanolicus
1.1.99.37 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

EC Number Metals/Ions Comment Organism Structure
1.1.1.244 Mg2+ bound to enzyme, essential for binding of NAD+ Bacillus methanolicus
1.1.1.244 Zn2+ bound to enzyme Bacillus methanolicus
1.1.99.37 Mg2+
-
 Bacillus methanolicus
1.1.99.37 Zn2+
-
 Bacillus methanolicus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.244 43000
-
 10 * 43000, SDS-PAGE Bacillus methanolicus
1.1.99.37 43000
-
 10 * 43000, SDS-PAGE Bacillus methanolicus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.244 methanol + NAD+ Bacillus methanolicus
-
 formaldehyde + NADH + H+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.244 Bacillus methanolicus
-
-
-
1.1.99.37 Bacillus methanolicus
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.244 wild type and mutant enzymes Bacillus methanolicus
1.1.99.37 recombinant enzyme Bacillus methanolicus

Reaction

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.99.37 0.413
-
 pH 9.5, 50°C Bacillus methanolicus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.244 methanol + NAD+
-
 Bacillus methanolicus formaldehyde + NADH + H+
-
 ?
1.1.99.37 formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
-
 Bacillus methanolicus methanol + N,N-dimethyl-4-nitrosoaniline
-
 r
1.1.99.37 methanol + N,N-dimethyl-4-nitrosoaniline
-
 Bacillus methanolicus formaldehyde + 4-(hydroxylamino)-N,N-dimethylaniline
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.244 decamer 10 * 43000, SDS-PAGE Bacillus methanolicus
1.1.99.37 decamer 10 * 43000, SDS-PAGE Bacillus methanolicus

Synonyms

EC Number Synonyms Comment Organism
1.1.1.244 More not identical with methanol dehydrogenase which uses pyrroloquinoline quinone as prosthetic group, EC 1.1.99.8 Bacillus methanolicus

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.244 NAD+ tightly but not covalentely bound to enzyme, a second NAD+ is required for the regeneration of bound NADH Bacillus methanolicus
1.1.99.37 NAD+
-
 Bacillus methanolicus
1.1.99.37 NADH
-
 Bacillus methanolicus