EC Number | Activating Compound | Comment | Organism | Structure |
---|---|---|---|---|
4.1.3.24 | acetate | growth of the mcl1::kan mutant on acetate is not rescued by the addition of glyoxylate, the mutant growth rates are comparable to those of the wild type on substrates downstream of the proposed metabolic block, i.e., malate and propionate plus bicarbonate | Cereibacter sphaeroides |
EC Number | Cloned (Comment) | Organism |
---|---|---|
3.1.2.30 | expression in Escherichia coli | Cereibacter sphaeroides |
4.1.3.24 | into pET16b and expressed as an N-terminal deca-His tag fusion protein in Escherichia coli BL21(DE3) cells | Cereibacter sphaeroides |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
3.1.2.30 | EDTA | reversible inhibition | Cereibacter sphaeroides | |
4.1.3.24 | Ca2+ | 14% specific activity at 10 mM | Cereibacter sphaeroides | |
4.1.3.24 | Co2+ | 13% specific activity at 10 mM | Cereibacter sphaeroides | |
4.1.3.24 | EDTA | 1% specific activity at 10 mM | Cereibacter sphaeroides | |
4.1.3.24 | Ni2+ | 6% specific activity at 10 mM | Cereibacter sphaeroides | |
4.1.3.24 | succinate | activity is downregulated 10fold | Cereibacter sphaeroides |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.1.2.30 | 0.09 | - |
(S)-malyl-CoA | pH 7.5, 30°C, in the presence of divalent cations (Mg2+ or Mn2+) | Cereibacter sphaeroides | |
4.1.3.24 | 0.01 | - |
beta-methylmalyl-CoA | at 30°C | Cereibacter sphaeroides | |
4.1.3.24 | 0.02 | - |
(3S)-malyl-CoA | at 30°C | Cereibacter sphaeroides | |
4.1.3.24 | 0.1 | - |
acetyl-CoA | at 30°C | Cereibacter sphaeroides | |
4.1.3.24 | 0.2 | - |
propionyl-CoA | at 30°C | Cereibacter sphaeroides | |
4.1.3.24 | 3.1 | - |
glyoxylate | condensation of acetyl-CoA and glyoxylate, at 30°C | Cereibacter sphaeroides | |
4.1.3.24 | 4.1 | - |
glyoxylate | condensation of propionyl-CoA and glyoxylate, at 30°C | Cereibacter sphaeroides |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
3.1.2.30 | Mg2+ | 10 mM, stimulated by Mg2+ or Mn2+ | Cereibacter sphaeroides | |
3.1.2.30 | Mn2+ | 10 mM, stimulated by Mg2+ or Mn2+ | Cereibacter sphaeroides | |
4.1.3.24 | Mg2+ | 73% specific activity at 10 mM | Cereibacter sphaeroides | |
4.1.3.24 | Mn2+ | 100% specific activity at 10 mM | Cereibacter sphaeroides |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
3.1.2.30 | 33400 | - |
2 * or 3 * x * 33400, calculated from sequence | Cereibacter sphaeroides |
3.1.2.30 | 80000 | - |
gel filtration | Cereibacter sphaeroides |
4.1.3.24 | 36800 | - |
4 * 36800, by sequence analysis | Cereibacter sphaeroides |
4.1.3.24 | 200000 | - |
gel filtration | Cereibacter sphaeroides |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.1.2.30 | (S)-malyl-CoA + H2O | Cereibacter sphaeroides | the enzyme is involved in the ethylmalonyl-CoA pathway for acetyl-CoA assimilation | (S)-malate + CoA | - |
? | |
3.1.2.30 | (S)-malyl-CoA + H2O | Cereibacter sphaeroides DSM 158 | the enzyme is involved in the ethylmalonyl-CoA pathway for acetyl-CoA assimilation | (S)-malate + CoA | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.1.2.30 | Cereibacter sphaeroides | D3JV05 | - |
- |
3.1.2.30 | Cereibacter sphaeroides DSM 158 | D3JV05 | - |
- |
4.1.3.24 | Cereibacter sphaeroides | Q3J5L6 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.1.2.30 | recombinant His-tagged proteins | Cereibacter sphaeroides |
4.1.3.24 | by affinity chromatography | Cereibacter sphaeroides |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
3.1.2.30 | 200 | - |
thioester cleavage, pH 7.5, 30°C | Cereibacter sphaeroides |
4.1.3.24 | 4.1 | - |
cleavage of (3S)-malyl-CoA | Cereibacter sphaeroides |
4.1.3.24 | 4.5 | - |
cleavage of beta-methylmalyl-CoA | Cereibacter sphaeroides |
4.1.3.24 | 14 | - |
condensation of acetyl-CoA and glyoxylate | Cereibacter sphaeroides |
4.1.3.24 | 20 | - |
condensation of propionyl-CoA and glyoxylate | Cereibacter sphaeroides |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.1.2.30 | (S)-malyl-CoA + H2O | the enzyme is involved in the ethylmalonyl-CoA pathway for acetyl-CoA assimilation | Cereibacter sphaeroides | (S)-malate + CoA | - |
? | |
3.1.2.30 | (S)-malyl-CoA + H2O | (S)-malyl-CoA i.e. (3S)-3-carboxy-3-hydroxypropanoyl-CoA, the enzyme has no activity with (2R,3S)-2-methylmalyl-CoA (cf. EC 4.1.3.24, malyl-CoA lyase) or other CoA esters | Cereibacter sphaeroides | (S)-malate + CoA | - |
? | |
3.1.2.30 | (S)-malyl-CoA + H2O | the enzyme is involved in the ethylmalonyl-CoA pathway for acetyl-CoA assimilation | Cereibacter sphaeroides DSM 158 | (S)-malate + CoA | - |
? | |
3.1.2.30 | (S)-malyl-CoA + H2O | (S)-malyl-CoA i.e. (3S)-3-carboxy-3-hydroxypropanoyl-CoA, the enzyme has no activity with (2R,3S)-2-methylmalyl-CoA (cf. EC 4.1.3.24, malyl-CoA lyase) or other CoA esters | Cereibacter sphaeroides DSM 158 | (S)-malate + CoA | - |
? | |
4.1.3.24 | (3S)-malyl-CoA | - |
Cereibacter sphaeroides | acetyl-CoA + glyoxylate | - |
r | |
4.1.3.24 | beta-methylmalyl-CoA | - |
Cereibacter sphaeroides | glyoxylate + propionyl-CoA | - |
r |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
3.1.2.30 | ? | 2 * or 3 * x * 33400, calculated from sequence | Cereibacter sphaeroides |
4.1.3.24 | homohexamer | 4 * 36800, by sequence analysis | Cereibacter sphaeroides |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.1.2.30 | Mcl2 | - |
Cereibacter sphaeroides |
4.1.3.24 | (3S)-malyl-CoA lyase | - |
Cereibacter sphaeroides |
4.1.3.24 | (3S)-malyl-coenzyme A lyase | - |
Cereibacter sphaeroides |
4.1.3.24 | malyl-CoA lyase | - |
Cereibacter sphaeroides |
4.1.3.24 | Mcl1 | - |
Cereibacter sphaeroides |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.1.2.30 | 30 | - |
assay at | Cereibacter sphaeroides |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.1.2.30 | 110 | - |
(S)-malyl-CoA | pH 7.5, 30°C | Cereibacter sphaeroides | |
4.1.3.24 | 2.5 | - |
(3S)-malyl-CoA | at 30°C | Cereibacter sphaeroides | |
4.1.3.24 | 2.8 | - |
beta-methylmalyl-CoA | at 30°C | Cereibacter sphaeroides | |
4.1.3.24 | 8.6 | - |
acetyl-CoA | at 30°C | Cereibacter sphaeroides | |
4.1.3.24 | 12 | - |
propionyl-CoA | at 30°C | Cereibacter sphaeroides |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
3.1.2.30 | 7.5 | - |
assay at | Cereibacter sphaeroides |
EC Number | General Information | Comment | Organism |
---|---|---|---|
3.1.2.30 | physiological function | the enzyme is involved in the ethylmalonyl-CoA pathway for acetyl-CoA assimilation | Cereibacter sphaeroides |
4.1.3.24 | metabolism | Mcl1 and Mcl2 together catalyze the apparent malate synthase activity during acetyl-CoA assimilation via the ethylmalonyl-CoA pathway. Mcl1 is a true (3S)-malyl-CoA lyase, catalyzing the Claisen condensation of acetyl-CoA with glyoxylate. Activity of the mcl1::kan mutant is not detected in cell extracts grown on succinate plus acetate. Apparent malate synthase activity is very low in cell extracts of the mcl1::kan and mcl2::kan mutants when grown on acetate or acetate plus succinate | Cereibacter sphaeroides |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.1.2.30 | 1200 | - |
(S)-malyl-CoA | pH 7.5, 30°C | Cereibacter sphaeroides |