BRENDA - Enzyme Database

Methanol:coenzyme M methyltransferase from Methanosarcina barkeri - substitution of the corrinoid harbouring subunit MtaC by free cob(I)alamin

Sauer, K.; Thauer, R.K.; Eur. J. Biochem. 261, 674-681 (1999)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
2.1.1.246
methanol
MtaB plus methanol positively affect the catalytic efficiency of MtaA. activation of MtaA by MtaB is methanol-dependent. Methylation of cob(I)inamide with methanol is dependent on imidazole but not on the demethylation of methylcob(III)inamide with coenzyme M. The demethylation reaction is even inhibited by imidazole
Methanosarcina barkeri
2.1.1.246
MtaB
MtaB plus methanol positively affect the catalytic efficiency of MtaA. activation of MtaA by MtaB is methanol-dependent. Methylation of cob(I)inamide with methanol is dependent on imidazole but not on the demethylation of methylcob(III)inamide with coenzyme M. The demethylation reaction is even inhibited by imidazole
Methanosarcina barkeri
2.1.1.246
Ti(III) citrate
increases the specific activity of MtaA by 60% and decreases the apparent Km for methylcob(III)-alamin from 0.003 mM to below 0.001 mM
Methanosarcina barkeri
Cloned(Commentary)
EC Number
Commentary
Organism
2.1.1.246
gene mtaA, expression of His-tagged MtaA in Escherichia coli strain M15
Methanosarcina barkeri
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.1.1.246
imidazole
the demethylation of cob(I)inamide reaction is inhibited by imidazole. Imidazole does not inhibit methyltransfer from methylcob(III)alamin to coenzyme M at 10 mM
Methanosarcina barkeri
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.1.1.246
additional information
-
additional information
MtaA kinetics, overview. Demethylation of methylcob(III)alamin catalysed by MtaA alone exhibit apparent Km for cob(I)alamin and methylcob(III)alamin of above 1 mm
Methanosarcina barkeri
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.1.1.246
Co2+
Zn2+ can be substituted by Co2+
Methanosarcina barkeri
2.1.1.246
Zn2+
Mta contains 1 mol Zn2+ per mol of enzyme, Zn2+ can be substituted by Co2+
Methanosarcina barkeri
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.1.1.246
a [methyl-Co(III) methanol-specific corrinoid protein] + coenzyme M
Methanosarcina barkeri
-
methyl-CoM + a [Co(I) methanol-specific corrinoid protein]
-
-
r
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.1.1.246
Methanosarcina barkeri
-
gene mtaA
-
Purification (Commentary)
EC Number
Commentary
Organism
2.1.1.246
recombinant His-tagged MtaA from Escherichia coli strain M15 by nickel affinity and anion exchange chromatography to homogeneity
Methanosarcina barkeri
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
2.1.1.246
0.2
-
MtaA with substrate methylcob(III)-alamin, pH 7.0, 37C
Methanosarcina barkeri
2.1.1.246
8
-
MtaA with substrate methylcob(III)inamide, 50 mM methylcob(III)inamide as substrate show an activity of 8 U/mg, approximately 40fold higher than with 50 mM methylcob(III)-alamin, pH 7.0, 37C
Methanosarcina barkeri
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.246
a [methyl-Co(III) methanol-specific corrinoid protein] + coenzyme M
-
717561
Methanosarcina barkeri
methyl-CoM + a [Co(I) methanol-specific corrinoid protein]
-
-
-
r
2.1.1.246
additional information
in the assay for methanol:coenzyme M methyltransferase activity cob(I)alamin can be substituted by cob(I)inamide which is devoid of the nucleotide loopmethylation of cob(I)inamide with methanol is dependent on imidazole but not on the demethylation of methylcob(III)inamide with coenzyme M
717561
Methanosarcina barkeri
?
-
-
-
-
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
2.1.1.246
37
-
assay at
Methanosarcina barkeri
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.1.1.246
7
-
assay at
Methanosarcina barkeri
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
2.1.1.246
methanol
MtaB plus methanol positively affect the catalytic efficiency of MtaA. activation of MtaA by MtaB is methanol-dependent. Methylation of cob(I)inamide with methanol is dependent on imidazole but not on the demethylation of methylcob(III)inamide with coenzyme M. The demethylation reaction is even inhibited by imidazole
Methanosarcina barkeri
2.1.1.246
MtaB
MtaB plus methanol positively affect the catalytic efficiency of MtaA. activation of MtaA by MtaB is methanol-dependent. Methylation of cob(I)inamide with methanol is dependent on imidazole but not on the demethylation of methylcob(III)inamide with coenzyme M. The demethylation reaction is even inhibited by imidazole
Methanosarcina barkeri
2.1.1.246
Ti(III) citrate
increases the specific activity of MtaA by 60% and decreases the apparent Km for methylcob(III)-alamin from 0.003 mM to below 0.001 mM
Methanosarcina barkeri
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.1.246
gene mtaA, expression of His-tagged MtaA in Escherichia coli strain M15
Methanosarcina barkeri
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.1.1.246
imidazole
the demethylation of cob(I)inamide reaction is inhibited by imidazole. Imidazole does not inhibit methyltransfer from methylcob(III)alamin to coenzyme M at 10 mM
Methanosarcina barkeri
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.1.1.246
additional information
-
additional information
MtaA kinetics, overview. Demethylation of methylcob(III)alamin catalysed by MtaA alone exhibit apparent Km for cob(I)alamin and methylcob(III)alamin of above 1 mm
Methanosarcina barkeri
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.1.1.246
Co2+
Zn2+ can be substituted by Co2+
Methanosarcina barkeri
2.1.1.246
Zn2+
Mta contains 1 mol Zn2+ per mol of enzyme, Zn2+ can be substituted by Co2+
Methanosarcina barkeri
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.1.1.246
a [methyl-Co(III) methanol-specific corrinoid protein] + coenzyme M
Methanosarcina barkeri
-
methyl-CoM + a [Co(I) methanol-specific corrinoid protein]
-
-
r
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.1.1.246
recombinant His-tagged MtaA from Escherichia coli strain M15 by nickel affinity and anion exchange chromatography to homogeneity
Methanosarcina barkeri
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
2.1.1.246
0.2
-
MtaA with substrate methylcob(III)-alamin, pH 7.0, 37C
Methanosarcina barkeri
2.1.1.246
8
-
MtaA with substrate methylcob(III)inamide, 50 mM methylcob(III)inamide as substrate show an activity of 8 U/mg, approximately 40fold higher than with 50 mM methylcob(III)-alamin, pH 7.0, 37C
Methanosarcina barkeri
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.1.1.246
a [methyl-Co(III) methanol-specific corrinoid protein] + coenzyme M
-
717561
Methanosarcina barkeri
methyl-CoM + a [Co(I) methanol-specific corrinoid protein]
-
-
-
r
2.1.1.246
additional information
in the assay for methanol:coenzyme M methyltransferase activity cob(I)alamin can be substituted by cob(I)inamide which is devoid of the nucleotide loopmethylation of cob(I)inamide with methanol is dependent on imidazole but not on the demethylation of methylcob(III)inamide with coenzyme M
717561
Methanosarcina barkeri
?
-
-
-
-
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
2.1.1.246
37
-
assay at
Methanosarcina barkeri
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.1.1.246
7
-
assay at
Methanosarcina barkeri
General Information
EC Number
General Information
Commentary
Organism
2.1.1.246
metabolism
methyl-coenzyme M formation from coenzyme M and methanol in Methanosarcina barkeri is catalysed by an enzyme system composed of three polypeptides MtaA, MtaB and MtaC, the latter of which harbours a corrinoid prosthetic group. We report here that MtaC can be substituted by free cob(I)alamin which is methylated with methanol in an MtaB-catalysed reaction and demethylated with coenzyme M in an MtaA-catalysed reaction
Methanosarcina barkeri
2.1.1.246
physiological function
a positive effect of MtaA on the catalytic efficiency of MtaB is specific for MtaA. In the absence of MtaA no effect is observed, while in the presence of MtaA the formation of methylcob(III)alamin from methanol and cob(I)alamin is apparently inhibited by coenzyme M, probably because under these conditions MtaA actively catalyses the demethylation of methylcob(III)alamin. MtaB plus methanol positively affect the catalytic efficiency of MtaA
Methanosarcina barkeri
General Information (protein specific)
EC Number
General Information
Commentary
Organism
2.1.1.246
metabolism
methyl-coenzyme M formation from coenzyme M and methanol in Methanosarcina barkeri is catalysed by an enzyme system composed of three polypeptides MtaA, MtaB and MtaC, the latter of which harbours a corrinoid prosthetic group. We report here that MtaC can be substituted by free cob(I)alamin which is methylated with methanol in an MtaB-catalysed reaction and demethylated with coenzyme M in an MtaA-catalysed reaction
Methanosarcina barkeri
2.1.1.246
physiological function
a positive effect of MtaA on the catalytic efficiency of MtaB is specific for MtaA. In the absence of MtaA no effect is observed, while in the presence of MtaA the formation of methylcob(III)alamin from methanol and cob(I)alamin is apparently inhibited by coenzyme M, probably because under these conditions MtaA actively catalyses the demethylation of methylcob(III)alamin. MtaB plus methanol positively affect the catalytic efficiency of MtaA
Methanosarcina barkeri