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Literature summary for 4.2.1.150 extracted from
- Identification and characterization of a highly thermostable crotonase from Meiothermus ruber (2015), J. Mol. Catal. B, 112, 40-44 .
No PubMed abstract available
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene Mrub_2284, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of C-terminally His-tagged enzyme in Escherichia coli strain Rosetta (DE3), subcloning in Escherichia coli strain XL-1 Blue | Meiothermus ruber |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| butanol | 50% inhibition at 2% v/v, 80% at 8% v/v | Meiothermus ruber |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.03 | - |
crotonyl-CoA | pH 7.0, 50°C, recombinant His-tagged enzyme | Meiothermus ruber | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 140000 | - |
native PAGE, recombinant enzyme | Meiothermus ruber |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| crotonyl-CoA + H2O | Meiothermus ruber | - |
3-hydroxybutyryl-CoA | - |
r | |
| crotonyl-CoA + H2O | Meiothermus ruber DSM 1279 | - |
3-hydroxybutyryl-CoA | - |
r | |
Organic Solvent Stability
| Organic Solvent | Comment | Organism |
|---|---|---|
| Butanol | inhibits the enzyme activity, 45% inhibition at 2% v/v, 80% at 8% v/v | Meiothermus ruber |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Meiothermus ruber | D3PLE5 | - |
- |
| Meiothermus ruber DSM 1279 | D3PLE5 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme from Escherichia coli strain Rosetta (DE3) by nickel affinity chromatography and desalting gel filtration | Meiothermus ruber |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| a short-chain (3S)-3-hydroxyacyl-CoA = a short-chain trans-2-enoyl-CoA + H2O | the Mr-Crt catalytic mechanism constitutes a concerted attack by the two glutamate residues. While Glu143 of Mr-Crt protonates the substrate, Glu123 abstracts a proton from a bound water molecule. The Gly120 activates the substrate by a hydrogen bond to the oxygen of the enoyl moiety of the CoA ester. The binding pocket for the CoA moiety is formed by characteristic hydrophobic amino acids and lysine residues | Meiothermus ruber |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| crotonyl-CoA + H2O | - |
Meiothermus ruber | 3-hydroxybutyryl-CoA | - |
r | |
| crotonyl-CoA + H2O | - |
Meiothermus ruber DSM 1279 | 3-hydroxybutyryl-CoA | - |
r | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homotetramer | 4 * 30000, about, sequence calculation | Meiothermus ruber |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| crotonase | - |
Meiothermus ruber |
| Mr-Crt | - |
Meiothermus ruber |
| Mrub_2284 | - |
Meiothermus ruber |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 55 | - |
- |
Meiothermus ruber |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | 78 | activity range, 20% of maximal activity at 30°C, 60% at 75°C, over 80% at 50-70°C, and maximal activity at 55°C | Meiothermus ruber |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 50 | - |
the purified recombinant His-tagged enzyme Mr-Crt displays an extended half-life of over 1 month, IT50: 743 h | Meiothermus ruber |
| 50 | 70 | purified recombinant His-tagged enzyme, 80% activity remaining after 5 min | Meiothermus ruber |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 14.9 | - |
crotonyl-CoA | pH 7.0, 50°C, recombinant His-tagged enzyme | Meiothermus ruber | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
- |
Meiothermus ruber |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6.5 | 7.5 | 60% of maximal activity is observed at pH 6.5 and pH 7.5 | Meiothermus ruber |
General Information
| General Information | Comment | Organism |
|---|---|---|
| additional information | The Mr-Crt catalytic triad is formed by residues Gly120, Glu123 and Glu143 | Meiothermus ruber |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 496.7 | - |
crotonyl-CoA | pH 7.0, 50°C, recombinant His-tagged enzyme | Meiothermus ruber | |
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