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- Epimerase (Msed_0639) and mutase (Msed_0638 and Msed_2055) convert (S)-methylmalonyl-coenzyme A (CoA) to succinyl-CoA in the Metallosphaera sedula 3-hydroxypropionate/4-hydroxybutyrate cycle (2012), Appl. Environ. Microbiol., 78, 6194-6202.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 5.1.99.1 | expression in Escherichia coli | Metallosphaera sedula |
| 5.1.99.1 | gene Msed_0639, DNA and amino acid sequence determination, phylogenetic tree, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain Rosetta 2 (DE3) | Metallosphaera sedula |
| 5.4.99.2 | expression in Escherichia coli | Metallosphaera sedula |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.1.99.1 | Co2+ | activates | Metallosphaera sedula |  |
| 5.1.99.1 | Co2+ | the enzyme is activated by divalent cations (Ni2+, Co2+, and Mg2+) | Metallosphaera sedula | |
| 5.1.99.1 | Mg2+ | activates | Metallosphaera sedula | |
| 5.1.99.1 | Mg2+ | the enzyme is activated by divalent cations (Ni2+, Co2+, and Mg2+) | Metallosphaera sedula | |
| 5.1.99.1 | Ni2+ | activates | Metallosphaera sedula | |
| 5.1.99.1 | Ni2+ | the enzyme is activated by divalent cations (Ni2+, Co2+, and Mg2+) | Metallosphaera sedula | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 5.1.99.1 | 20000 | - |
2 * 20000, SDS-PAGE | Metallosphaera sedula |
| 5.1.99.1 | 20000 | - |
2 * 20000, recombinant His-tagged enzyme, SDS-PAGE | Metallosphaera sedula |
| 5.1.99.1 | 32100 | - |
gel filtration | Metallosphaera sedula |
| 5.1.99.1 | 32100 | - |
recombinant His-tagged enzyme, gel filtration | Metallosphaera sedula |
| 5.4.99.2 | 17000 | - |
2 * 17000 + 2 * 70000, SDS-PAGE | Metallosphaera sedula |
| 5.4.99.2 | 70000 | - |
2 * 17000 + 2 * 70000, SDS-PAGE | Metallosphaera sedula |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.1.99.1 | (S)-methylmalonyl-CoA | Metallosphaera sedula | - |
(R)-methylmalonyl-CoA | - |
? | |
| 5.1.99.1 | (S)-methylmalonyl-CoA | Metallosphaera sedula | the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway | (R)-methylmalonyl-CoA | - |
? | |
| 5.1.99.1 | (S)-methylmalonyl-CoA | Metallosphaera sedula DSM 5348 | - |
(R)-methylmalonyl-CoA | - |
? | |
| 5.1.99.1 | (S)-methylmalonyl-CoA | Metallosphaera sedula DSM 5348 | the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway | (R)-methylmalonyl-CoA | - |
? | |
| 5.4.99.2 | (R)-methylmalonyl-CoA | Metallosphaera sedula | the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway | succinyl-CoA | - |
r | |
| 5.4.99.2 | (R)-methylmalonyl-CoA | Metallosphaera sedula DSM 5348 | the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway | succinyl-CoA | - |
r | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 5.1.99.1 | Metallosphaera sedula | A4YEG2 | - |
- |
| 5.1.99.1 | Metallosphaera sedula DSM 5348 | A4YEG2 | - |
- |
| 5.4.99.2 | Metallosphaera sedula | A4YEG1 and A4YIE3 | A4YEG1: catalytic subunit of methylmalonyl-CoA mutase, A4YIE3: coenzyme B12-binding subunit of methylmalonyl-CoA mutase | - |
| 5.4.99.2 | Metallosphaera sedula DSM 5348 | A4YEG1 and A4YIE3 | A4YEG1: catalytic subunit of methylmalonyl-CoA mutase, A4YIE3: coenzyme B12-binding subunit of methylmalonyl-CoA mutase | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 5.1.99.1 | - |
Metallosphaera sedula |
| 5.1.99.1 | recombinant N-terminally His-tagged enzyme from Escherichia coli strain Rosetta 2 (DE3) by nickel affinity chromatography | Metallosphaera sedula |
| 5.4.99.2 | - |
Metallosphaera sedula |
Source Tissue
| EC Number | Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|---|
| 5.1.99.1 | additional information | transcriptomic analyses of Metallosphaera sedula grown under autotrophic versus heterotrophic conditions, overview. MCE expression is significantly upregulated under autotrophic compared to heterotrophic growth conditions | Metallosphaera sedula | - |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 5.1.99.1 | 218 | - |
75°C, pH not specified in the publication | Metallosphaera sedula |
| 5.1.99.1 | 218 | - |
purified recombinant enzyme, pH 7.0, 75°C | Metallosphaera sedula |
| 5.4.99.2 | 2.2 | - |
75°C, pH not specified in the publication | Metallosphaera sedula |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.1.99.1 | (S)-methylmalonyl-CoA | - |
Metallosphaera sedula | (R)-methylmalonyl-CoA | - |
? | |
| 5.1.99.1 | (S)-methylmalonyl-CoA | the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway | Metallosphaera sedula | (R)-methylmalonyl-CoA | - |
? | |
| 5.1.99.1 | (S)-methylmalonyl-CoA | - |
Metallosphaera sedula DSM 5348 | (R)-methylmalonyl-CoA | - |
? | |
| 5.1.99.1 | (S)-methylmalonyl-CoA | the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway | Metallosphaera sedula DSM 5348 | (R)-methylmalonyl-CoA | - |
? | |
| 5.4.99.2 | (R)-methylmalonyl-CoA | - |
Metallosphaera sedula | succinyl-CoA | - |
r | |
| 5.4.99.2 | (R)-methylmalonyl-CoA | the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway | Metallosphaera sedula | succinyl-CoA | - |
r | |
| 5.4.99.2 | (R)-methylmalonyl-CoA | - |
Metallosphaera sedula DSM 5348 | succinyl-CoA | - |
r | |
| 5.4.99.2 | (R)-methylmalonyl-CoA | the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway | Metallosphaera sedula DSM 5348 | succinyl-CoA | - |
r | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 5.1.99.1 | homodimer | 2 * 20000, SDS-PAGE | Metallosphaera sedula |
| 5.1.99.1 | homodimer | 2 * 20000, recombinant His-tagged enzyme, SDS-PAGE | Metallosphaera sedula |
| 5.1.99.1 | More | the putative three-dimensional structure of the enzyme is modeled using hypothetical protein BH1468, PDB ID 3OA4 | Metallosphaera sedula |
| 5.4.99.2 | heterotetramer | 2 * 17000 + 2 * 70000, SDS-PAGE | Metallosphaera sedula |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 5.1.99.1 | MCE | - |
Metallosphaera sedula |
| 5.1.99.1 | methylmalonyl-CoA epimerase | - |
Metallosphaera sedula |
| 5.1.99.1 | methylmalonyl-coenzyme A epimerase | - |
Metallosphaera sedula |
| 5.1.99.1 | Msed_0639 | locus name | Metallosphaera sedula |
| 5.4.99.2 | MCM | - |
Metallosphaera sedula |
| 5.4.99.2 | Msed_0638 | locus name, catalytic subunit of methylmalonyl-CoA mutase | Metallosphaera sedula |
| 5.4.99.2 | Msed_2055 | locus name, coenzyme B12-binding subunit of methylmalonyl-CoA mutase | Metallosphaera sedula |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 5.1.99.1 | 73 | - |
- |
Metallosphaera sedula |
| 5.1.99.1 | 75 | - |
assay at | Metallosphaera sedula |
| 5.4.99.2 | 75 | - |
assay at | Metallosphaera sedula |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 5.1.99.1 | 7 | - |
assay at | Metallosphaera sedula |
Expression
| EC Number | Organism | Comment | Expression |
|---|---|---|---|
| 5.1.99.1 | Metallosphaera sedula | expression of the gene is significantly upregulated under autotrophic compared to heterotrophic growth conditions, implying a role in CO2 fixation | up |
| 5.1.99.1 | Metallosphaera sedula | the enzyme expression is significantly upregulated under autotrophic compared to heterotrophic growth conditions | up |
| 5.4.99.2 | Metallosphaera sedula | expression of the gene is significantly upregulated under autotrophic compared to heterotrophic growth conditions, implying a role in CO2 fixation | up |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 5.1.99.1 | physiological function | methylmalonyl-CoA epimerase and methylmalonyl-CoA mutase are responsible for producing succinyl-CoA from (S)-methylmalonyl-CoA. Succinyl-CoA is an intermediate connecting the 3-hydroxypropionate/4-hydroxybutyrate cycle and general carbon metabolism. Methylmalonyl-CoA epimerase and methylmalonyl-CoA mutase may also be involved in degradation of propionate and branched-chain amino acids (valine, isoleucine, methionine). The enzyme has a role in CO2 fixation | Metallosphaera sedula |
| 5.1.99.1 | physiological function | the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway | Metallosphaera sedula |
| 5.4.99.2 | physiological function | the enzyme is involved in the 3-hydroxypropionate/4-hydroxybutyrate carbon fixation pathway | Metallosphaera sedula |
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