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Literature summary for 1.1.1.244 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

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

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli DH5alpha Bacillus methanolicus

Protein Variants

Protein Variants Comment Organism
D100N strongly reduced NAD+-binding, no activity Bacillus methanolicus
D88N only minor effects on activity Bacillus methanolicus
G13A only minor effects on activity Bacillus methanolicus
G15A only minor effects on activity Bacillus methanolicus
G95A impaired cofactor binding, low acitivity Bacillus methanolicus
K103R strongly reduced NAD+-binding, no activity Bacillus methanolicus
S97G impaired cofactor binding, much higher acitivity than the wild type enzyme, no activation with ACT protein Bacillus methanolicus
S97T impaired cofactor binding, much higher acitivity than the wild type enzyme, no activation with ACT protein Bacillus methanolicus

KM Value [mM]

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

Metals/Ions

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Bacillus methanolicus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
wild type and mutant enzymes Bacillus methanolicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
methanol + NAD+
-
Bacillus methanolicus formaldehyde + NADH + H+
-
?

Subunits

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

Synonyms

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

Cofactor

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