BRENDA - Enzyme Database

Ethylmalonyl-CoA mutase from Rhodobacter sphaeroides defines a new subclade of coenzyme B12-dependent acyl-CoA mutases

Erb, T.J.; Retey, J.; Fuchs, G.; Alber, B.E.; J. Biol. Chem. 283, 32283-32293 (2008)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
5.1.99.1
expression in Eschserichia coli
Rhodobacter sphaeroides
5.4.99.63
expression in Escherichia coli
Rhodobacter sphaeroides
Crystallization (Commentary)
EC Number
Crystallization (Commentary)
Organism
5.4.99.63
computational modeling using the methylmalonyl-CoA mutase substrate complex from P. freudenreichii subsp. shermanii, Protein Data Bank code 4REQ as template
Rhodobacter sphaeroides
Engineering
EC Number
Protein Variants
Commentary
Organism
5.4.99.63
additional information
gene insertion mutant is unable to use acetate or acetoacetate as the sole carbon source. Growth on propionate/HCO3-, which requires methylmalonate-CoA mutase, is not affected. Mutant cells do not display ethylmalonyl-CoA converting activity but convert methylmalonyl-CoA to succinyl-CoA
Rhodobacter sphaeroides
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
5.1.99.1
EDTA
2 mM, complete loss of activity
Rhodobacter sphaeroides
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
5.1.99.1
0.04
-
(2S)-ethylmalonyl-CoA
-
Rhodobacter sphaeroides
5.1.99.1
0.08
-
(S)-methylmalonyl-CoA
-
Rhodobacter sphaeroides
5.4.99.63
0.06
-
(2R)-ethylmalonyl-CoA
pH 7.8, 30°C
Rhodobacter sphaeroides
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
5.1.99.1
Co2+
0.4 mM, 4fold increase in activity. Presence of Co2+ restores activity after incubation with EDTA
Rhodobacter sphaeroides
5.1.99.1
Mn2+
presence of Mn2+ restores activity after incubation with EDTA
Rhodobacter sphaeroides
5.1.99.1
additional information
no activation of enzyme with Mg2+, Ni2+
Rhodobacter sphaeroides
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
5.4.99.63
74000
-
2 * 75000, SDS-PAGE, 2 * 74000, calculated
Rhodobacter sphaeroides
5.4.99.63
75000
-
2 * 75000, SDS-PAGE, 2 * 74000, calculated
Rhodobacter sphaeroides
5.4.99.63
153000
-
gel filtration
Rhodobacter sphaeroides
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
5.1.99.1
additional information
Rhodobacter sphaeroides
enzyme catalyzes the transformation of ethylmalonyl-CoA to methylsuccinyl-CoA in combination with ethylmalonyl-CoA mutase, in a ethylmalonyl-CoA dependent acyl-CoA pathway for acetate assimilation
?
-
-
?
5.4.99.63
additional information
Rhodobacter sphaeroides
enzyme catalyzes the transformation of ethylmalonyl-CoA to methylsuccinyl-CoA in combination with ethylmalonyl-CoA racemase in a ethylmalonyl-CoA dependent acyl-CoA pathway for acetate assimilation
?
-
-
?
Organism
EC Number
Organism
UniProt
Commentary
Textmining
5.1.99.1
Rhodobacter sphaeroides
Q3IZP4
promiscuous ethylmalonyl-CoA/methylmalonyl-CoA epimerase
-
5.4.99.63
Rhodobacter sphaeroides
Q3IZ90
-
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
5.1.99.1
recombinant enzyme
Rhodobacter sphaeroides
5.4.99.63
recombinant enzyme. Purified recombinant Ecm is unable to form methylsuccinyl-CoA from ethylmalonyl-CoA when ethylmalonyl-CoA is produced by reductive carboxylation of crotonyl-CoA carboxylase/reductase. Addition of cell extract of the gene insertion mutant ecm::kan to the enzyme assay restores ethylmalonyl-CoA mutase activity
Rhodobacter sphaeroides
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
5.4.99.63
0.05
-
pH 7.8, 30°C
Rhodobacter sphaeroides
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
5.1.99.1
(2S)-ethylmalonyl-CoA
-
693174
Rhodobacter sphaeroides
(2R)-ethylmalonyl-CoA
-
-
-
?
5.1.99.1
(S)-methylmalonyl-CoA
-
693174
Rhodobacter sphaeroides
(R)-methylmalonyl-CoA
-
-
-
?
5.1.99.1
additional information
enzyme catalyzes the transformation of ethylmalonyl-CoA to methylsuccinyl-CoA in combination with ethylmalonyl-CoA mutase, in a ethylmalonyl-CoA dependent acyl-CoA pathway for acetate assimilation
693174
Rhodobacter sphaeroides
?
-
-
-
?
5.4.99.63
(2R)-ethylmalonyl-CoA
purified recombinant Ecm is unable to form methylsuccinyl-CoA from ethylmalonyl-CoA when ethylmalonyl-CoA is produced by reductive carboxylation of crotonyl-CoA carboxylase/reductase. Addition of cell extract of the gene insertion mutant ecm::kan to the enzyme assay restores ethylmalonyl-CoA mutase activity due to presence of ethylmalonyl-CoA epimerase
693174
Rhodobacter sphaeroides
(2S)-methylsuccinyl-CoA
-
-
-
?
5.4.99.63
(R)-2-methylmalonyl-CoA
0.2% of the activity with (2R)-ethylmalonyl-CoA
693174
Rhodobacter sphaeroides
succinyl-CoA
-
-
-
?
5.4.99.63
additional information
enzyme catalyzes the transformation of ethylmalonyl-CoA to methylsuccinyl-CoA in combination with ethylmalonyl-CoA racemase in a ethylmalonyl-CoA dependent acyl-CoA pathway for acetate assimilation
693174
Rhodobacter sphaeroides
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
5.4.99.63
dimer
2 * 75000, SDS-PAGE, 2 * 74000, calculated
Rhodobacter sphaeroides
Synonyms
EC Number
Synonyms
Commentary
Organism
5.4.99.63
Ecm
-
Rhodobacter sphaeroides
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
5.4.99.63
6.5
8
broad
Rhodobacter sphaeroides
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
5.4.99.63
vitamin B12
Km value 0.002 mM
Rhodobacter sphaeroides
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
5.1.99.1
expression in Eschserichia coli
Rhodobacter sphaeroides
5.4.99.63
expression in Escherichia coli
Rhodobacter sphaeroides
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
5.4.99.63
vitamin B12
Km value 0.002 mM
Rhodobacter sphaeroides
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
5.4.99.63
computational modeling using the methylmalonyl-CoA mutase substrate complex from P. freudenreichii subsp. shermanii, Protein Data Bank code 4REQ as template
Rhodobacter sphaeroides
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
5.4.99.63
additional information
gene insertion mutant is unable to use acetate or acetoacetate as the sole carbon source. Growth on propionate/HCO3-, which requires methylmalonate-CoA mutase, is not affected. Mutant cells do not display ethylmalonyl-CoA converting activity but convert methylmalonyl-CoA to succinyl-CoA
Rhodobacter sphaeroides
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
5.1.99.1
EDTA
2 mM, complete loss of activity
Rhodobacter sphaeroides
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
5.1.99.1
0.04
-
(2S)-ethylmalonyl-CoA
-
Rhodobacter sphaeroides
5.1.99.1
0.08
-
(S)-methylmalonyl-CoA
-
Rhodobacter sphaeroides
5.4.99.63
0.06
-
(2R)-ethylmalonyl-CoA
pH 7.8, 30°C
Rhodobacter sphaeroides
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
5.1.99.1
Co2+
0.4 mM, 4fold increase in activity. Presence of Co2+ restores activity after incubation with EDTA
Rhodobacter sphaeroides
5.1.99.1
Mn2+
presence of Mn2+ restores activity after incubation with EDTA
Rhodobacter sphaeroides
5.1.99.1
additional information
no activation of enzyme with Mg2+, Ni2+
Rhodobacter sphaeroides
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
5.4.99.63
74000
-
2 * 75000, SDS-PAGE, 2 * 74000, calculated
Rhodobacter sphaeroides
5.4.99.63
75000
-
2 * 75000, SDS-PAGE, 2 * 74000, calculated
Rhodobacter sphaeroides
5.4.99.63
153000
-
gel filtration
Rhodobacter sphaeroides
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
5.1.99.1
additional information
Rhodobacter sphaeroides
enzyme catalyzes the transformation of ethylmalonyl-CoA to methylsuccinyl-CoA in combination with ethylmalonyl-CoA mutase, in a ethylmalonyl-CoA dependent acyl-CoA pathway for acetate assimilation
?
-
-
?
5.4.99.63
additional information
Rhodobacter sphaeroides
enzyme catalyzes the transformation of ethylmalonyl-CoA to methylsuccinyl-CoA in combination with ethylmalonyl-CoA racemase in a ethylmalonyl-CoA dependent acyl-CoA pathway for acetate assimilation
?
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
5.1.99.1
recombinant enzyme
Rhodobacter sphaeroides
5.4.99.63
recombinant enzyme. Purified recombinant Ecm is unable to form methylsuccinyl-CoA from ethylmalonyl-CoA when ethylmalonyl-CoA is produced by reductive carboxylation of crotonyl-CoA carboxylase/reductase. Addition of cell extract of the gene insertion mutant ecm::kan to the enzyme assay restores ethylmalonyl-CoA mutase activity
Rhodobacter sphaeroides
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
5.4.99.63
0.05
-
pH 7.8, 30°C
Rhodobacter sphaeroides
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
5.1.99.1
(2S)-ethylmalonyl-CoA
-
693174
Rhodobacter sphaeroides
(2R)-ethylmalonyl-CoA
-
-
-
?
5.1.99.1
(S)-methylmalonyl-CoA
-
693174
Rhodobacter sphaeroides
(R)-methylmalonyl-CoA
-
-
-
?
5.1.99.1
additional information
enzyme catalyzes the transformation of ethylmalonyl-CoA to methylsuccinyl-CoA in combination with ethylmalonyl-CoA mutase, in a ethylmalonyl-CoA dependent acyl-CoA pathway for acetate assimilation
693174
Rhodobacter sphaeroides
?
-
-
-
?
5.4.99.63
(2R)-ethylmalonyl-CoA
purified recombinant Ecm is unable to form methylsuccinyl-CoA from ethylmalonyl-CoA when ethylmalonyl-CoA is produced by reductive carboxylation of crotonyl-CoA carboxylase/reductase. Addition of cell extract of the gene insertion mutant ecm::kan to the enzyme assay restores ethylmalonyl-CoA mutase activity due to presence of ethylmalonyl-CoA epimerase
693174
Rhodobacter sphaeroides
(2S)-methylsuccinyl-CoA
-
-
-
?
5.4.99.63
(R)-2-methylmalonyl-CoA
0.2% of the activity with (2R)-ethylmalonyl-CoA
693174
Rhodobacter sphaeroides
succinyl-CoA
-
-
-
?
5.4.99.63
additional information
enzyme catalyzes the transformation of ethylmalonyl-CoA to methylsuccinyl-CoA in combination with ethylmalonyl-CoA racemase in a ethylmalonyl-CoA dependent acyl-CoA pathway for acetate assimilation
693174
Rhodobacter sphaeroides
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
5.4.99.63
dimer
2 * 75000, SDS-PAGE, 2 * 74000, calculated
Rhodobacter sphaeroides
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
5.4.99.63
6.5
8
broad
Rhodobacter sphaeroides