EC Number | Cloned (Comment) | Organism |
---|---|---|
5.1.99.1 | expression in Eschserichia coli | Cereibacter sphaeroides |
5.4.99.63 | expression in Escherichia coli | Cereibacter sphaeroides |
EC Number | Crystallization (Comment) | 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 | Cereibacter sphaeroides |
EC Number | Protein Variants | Comment | 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 | Cereibacter sphaeroides |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
5.1.99.1 | EDTA | 2 mM, complete loss of activity | Cereibacter sphaeroides |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
5.1.99.1 | 0.04 | - |
(2S)-ethylmalonyl-CoA | - |
Cereibacter sphaeroides | |
5.1.99.1 | 0.08 | - |
(S)-methylmalonyl-CoA | - |
Cereibacter sphaeroides | |
5.4.99.63 | 0.06 | - |
(2R)-ethylmalonyl-CoA | pH 7.8, 30°C | Cereibacter sphaeroides |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
5.1.99.1 | Co2+ | 0.4 mM, 4fold increase in activity. Presence of Co2+ restores activity after incubation with EDTA | Cereibacter sphaeroides | |
5.1.99.1 | Mn2+ | presence of Mn2+ restores activity after incubation with EDTA | Cereibacter sphaeroides | |
5.1.99.1 | additional information | no activation of enzyme with Mg2+, Ni2+ | Cereibacter sphaeroides |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
5.4.99.63 | 74000 | - |
2 * 75000, SDS-PAGE, 2 * 74000, calculated | Cereibacter sphaeroides |
5.4.99.63 | 75000 | - |
2 * 75000, SDS-PAGE, 2 * 74000, calculated | Cereibacter sphaeroides |
5.4.99.63 | 153000 | - |
gel filtration | Cereibacter sphaeroides |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
5.1.99.1 | additional information | Cereibacter 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 | Cereibacter 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 | ? | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
5.1.99.1 | Cereibacter sphaeroides | Q3IZP4 | promiscuous ethylmalonyl-CoA/methylmalonyl-CoA epimerase | - |
5.4.99.63 | Cereibacter sphaeroides | Q3IZ90 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
5.1.99.1 | recombinant enzyme | Cereibacter 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 | Cereibacter sphaeroides |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
5.4.99.63 | 0.05 | - |
pH 7.8, 30°C | Cereibacter sphaeroides |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
5.1.99.1 | (2S)-ethylmalonyl-CoA | - |
Cereibacter sphaeroides | (2R)-ethylmalonyl-CoA | - |
? | |
5.1.99.1 | (S)-methylmalonyl-CoA | - |
Cereibacter 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 | Cereibacter 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 | Cereibacter sphaeroides | (2S)-methylsuccinyl-CoA | - |
? | |
5.4.99.63 | (R)-2-methylmalonyl-CoA | 0.2% of the activity with (2R)-ethylmalonyl-CoA | Cereibacter 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 | Cereibacter sphaeroides | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
5.4.99.63 | dimer | 2 * 75000, SDS-PAGE, 2 * 74000, calculated | Cereibacter sphaeroides |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
5.4.99.63 | Ecm | - |
Cereibacter sphaeroides |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
5.4.99.63 | 6.5 | 8 | broad | Cereibacter sphaeroides |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
5.4.99.63 | vitamin B12 | Km value 0.002 mM | Cereibacter sphaeroides |