Activating Compound | Comment | Organism | Structure |
---|---|---|---|
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 |
Cloned (Comment) | Organism |
---|---|
into pET16b and expressed as an N-terminal deca-His tag fusion protein in Escherichia coli BL21(DE3) cells | Cereibacter sphaeroides |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | 14% specific activity at 10 mM | Cereibacter sphaeroides | |
Co2+ | 13% specific activity at 10 mM | Cereibacter sphaeroides | |
EDTA | 1% specific activity at 10 mM | Cereibacter sphaeroides | |
Ni2+ | 6% specific activity at 10 mM | Cereibacter sphaeroides | |
succinate | activity is downregulated 10fold | Cereibacter sphaeroides |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.01 | - |
beta-methylmalyl-CoA | at 30°C | Cereibacter sphaeroides | |
0.02 | - |
(3S)-malyl-CoA | at 30°C | Cereibacter sphaeroides | |
0.1 | - |
acetyl-CoA | at 30°C | Cereibacter sphaeroides | |
0.2 | - |
propionyl-CoA | at 30°C | Cereibacter sphaeroides | |
3.1 | - |
glyoxylate | condensation of acetyl-CoA and glyoxylate, at 30°C | Cereibacter sphaeroides | |
4.1 | - |
glyoxylate | condensation of propionyl-CoA and glyoxylate, at 30°C | Cereibacter sphaeroides |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | 73% specific activity at 10 mM | Cereibacter sphaeroides | |
Mn2+ | 100% specific activity at 10 mM | Cereibacter sphaeroides |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
36800 | - |
4 * 36800, by sequence analysis | Cereibacter sphaeroides |
200000 | - |
gel filtration | Cereibacter sphaeroides |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Cereibacter sphaeroides | Q3J5L6 | - |
- |
Purification (Comment) | Organism |
---|---|
by affinity chromatography | Cereibacter sphaeroides |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
4.1 | - |
cleavage of (3S)-malyl-CoA | Cereibacter sphaeroides |
4.5 | - |
cleavage of beta-methylmalyl-CoA | Cereibacter sphaeroides |
14 | - |
condensation of acetyl-CoA and glyoxylate | Cereibacter sphaeroides |
20 | - |
condensation of propionyl-CoA and glyoxylate | Cereibacter sphaeroides |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(3S)-malyl-CoA | - |
Cereibacter sphaeroides | acetyl-CoA + glyoxylate | - |
r | |
beta-methylmalyl-CoA | - |
Cereibacter sphaeroides | glyoxylate + propionyl-CoA | - |
r |
Subunits | Comment | Organism |
---|---|---|
homohexamer | 4 * 36800, by sequence analysis | Cereibacter sphaeroides |
Synonyms | Comment | Organism |
---|---|---|
(3S)-malyl-CoA lyase | - |
Cereibacter sphaeroides |
(3S)-malyl-coenzyme A lyase | - |
Cereibacter sphaeroides |
malyl-CoA lyase | - |
Cereibacter sphaeroides |
Mcl1 | - |
Cereibacter sphaeroides |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.5 | - |
(3S)-malyl-CoA | at 30°C | Cereibacter sphaeroides | |
2.8 | - |
beta-methylmalyl-CoA | at 30°C | Cereibacter sphaeroides | |
8.6 | - |
acetyl-CoA | at 30°C | Cereibacter sphaeroides | |
12 | - |
propionyl-CoA | at 30°C | Cereibacter sphaeroides |
General Information | Comment | Organism |
---|---|---|
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 |